bromochloroacetic-acid and Mouth-Neoplasms

Papillary squamous cell carcinoma (SCC) (PSCC) of the oral mucosa is a relatively rare but distinct variant of SCC of head and neck. The objectives of this study were to describe the clinicopathologic and immunohistochemical features of a series of patients with oral PSCC and to review the literature on this topic. Retrospective review of patients with clinical and pathologic diagnosis of PSCC (n = 12) between 2000 and 2008 in our institution was conducted. The outcome analysis in a mean follow-up of 56 months (range, 24-131 months) was performed. These patients were 7 women and 5 men, and the mean age at diagnosis was 72.9 years (range, 53-83 years). The cheek and the gingiva were the predominant sites of involvement. At the end of follow-up, 4 patients were found to have local recurrence, and 3 were dead of disease. The estimated 3- and 5-year survival was 91.7% and 76.4% for the whole series, respectively. Histopathologically, the papillary pattern consisted of multiple, thin, delicate filiform, finger-like papillary projections with fibrovascular cores. Besides, the exophytic pattern consisted of the broad-based bulbous to "cauliflower-like" exophytic growth with rounded projections. Immunohistochemically, positivity for CKpan, CKhmw (high molecular weight), and p53, yet negativity for CK8, vimentin, desmin, smooth muscle actin, and S-100 was observed in PSCC. In conclusion, 2 specific histopathologic growth patterns of oral PSCC were identified to separate from conventional SCC. Patients with PSCC have a favorable outcome in relation to exophytic nature and limited invasion of the tumor.

Topics: Aged; Aged, 80 and over; Carcinoma, Papillary; Carcinoma, Squamous Cell; Cheek; Female; Gingiva; Humans; Kaplan-Meier Estimate; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Retrospective Studies; Survival Rate; Tumor Suppressor Protein p53

During the last decade, oral cytology has once again become the focus of scientific research. This new interest is due to the introduction of a cytobrush for cell collection as well as a computer-assisted analysis (Oral CDx). Although promising, the sensitivity and specificity of conventional oral brush cytology remains limited. To circumvent the problems and improve the accuracy, various adjunctive analytical methods have been attempted. DNA analysis, immunocytochemical and molecular analysis are suggested methodological cytology approaches to improve the validity of oral brush cytology. An increase in sensitivity (up to 100%) and specificity (up to 100%) of oral brush biopsy has been reported on localized pre-malignant and malignant lesions. Oral brush biopsy probably will not replace histopathology in the definitive diagnosis of oral cancer, but it might be valuable for the prevention of misdiagnosis of clinically doubtful oral lesions and for the monitoring of lesions that might proceed on to oral cancer.

Topics: Cell Shape; Cytodiagnosis; Histocytochemistry; Humans; Image Cytometry; Image Processing, Computer-Assisted; Keratins; Mouth Neoplasms; Neoplasm Proteins; Nucleolus Organizer Region; Ploidies; Protein Array Analysis; Sensitivity and Specificity

Thyroid carcinomas have been found incidentally in the cervical lymph nodes during surgery for head and neck squamous cell carcinoma (SCC). Such carcinomas have been considered a metastatic focus for malignant transformation of heterotopic thyroid tissue. We report on cases of so-called occult thyroid carcinoma in the cervical lymph nodes, and review the relevant literature.. We encountered 3 cases of incidental papillary carcinoma in the cervical lymph nodes of patients with oral SCC, and consequently reviewed 75 previously reported cases.. Among 148 patients with oral SCC who had undergone cervical lymph node dissection, 3 were diagnosed with occult thyroid carcinoma. Papillary carcinomas were found in 3, 10, and 3 lymph nodes in cases 1, 2, and 3, respectively. Computed tomography showed 2 tumor-like shadows and 1 calcified mass in the thyroid gland in cases 2 and 3, respectively. These shadows did not enlarge during the 3 to 5 years of observation, and all patients are alive, without any events related to the neck and thyroid gland. Among the reviewed cases, approximately two fifths were histopathologically or clinically free from thyroid carcinoma. Progressive thyroid carcinoma was not detected in any patient.. We propose the possibility that thyroid carcinoma in the cervical lymph nodes is not necessarily metastatic, but may occasionally arise from heterotopic thyroid tissue.

Topics: Aged; Carcinoma, Papillary; Carcinoma, Squamous Cell; Choristoma; Diagnosis, Differential; Histocytochemistry; Humans; Incidental Findings; Keratins; Lymph Nodes; Male; Middle Aged; Mouth Neoplasms; Neck; Neck Dissection; Thyroglobulin; Thyroid Gland; Thyroid Neoplasms

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy and is a major cause of cancer morbidity and mortality worldwide. Oral cancer is the most predominant malignancy in the Indian subcontinent due to the widespread habits of chewing tobacco and related products. Patients with oral tumours have a high risk of early locoregional relapse. Early detection of disease progression remains a challenging task mainly due to the lack of adequate early prognostic markers. CEA, SCC Ag, CA-125, serum cytokeratin (CK) fragments, Cyfra 21-1 (CK 19), TPS (CK 18), TPA (CK 8, 18, and 19) etc. are being used as serum markers for the prediction of prognosis of various malignancies. This review presents the available literature on serum CK markers in different malignancies evaluates their utility in the management of oral cancer, and identifies the lacunae which need to be addressed to develop sensitive and specific assays for early detection of recurrence, prognosis, and treatment monitoring.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; India; Keratins; Lymphatic Metastasis; Male; Mouth Neoplasms; Neoplasm Staging; Prognosis; Tobacco, Smokeless

Topics: Animals; Genetic Therapy; Herpesviridae Infections; Herpesvirus 1, Human; Humans; Keratins; Mouth Diseases; Mouth Neoplasms; Mutation; Periodontitis; Reactive Oxygen Species; Recurrence

Cytokeratins (CK) are being extensively used as diagnostic markers for various malignancies and other diseases, including human oral precancer and cancer, due to their tissue specific expression. CK are epithelia specific intermediate filament (IF) proteins, which are expressed in a differentiation dependent and tissue specific manner. There are about 30 polypeptides of CK expressed by different human epithelia. Each type of epithelium expresses about 4-6 polypeptides. CK polypeptides share many common epitopes, due to which the antibodies developed against CK tend to cross react. Therefore, a large number of monoclonal and polyclonal antibodies have been developed to distinguish among these proteins. Many of these antibodies are not only monospecific but are also epitope specific. These antibodies are being used in pathology laboratories for routine diagnosis using immunohistochemistry. A number of fixatives are used for fixation of tissue sections prior to the use of these antibodies. Sometimes, this leads in epitope masking. Hence, it becomes necessary to use a battery of monoclonal antibodies (MAb) for accurate diagnosis. Apart from the use of these antibodies in diagnostics, they are also being used in basic research for the study of CK function and their interactions with associated proteins and membrane proteins. In the present communication an effort has been made to make a comprehensive list of MAb to CK giving information like cross-reactivity, epitope specificity, various fixatives used, etc. along with the source of the antibodies, which will provide useful information to the users.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Databases, Factual; Epitopes; Humans; Immunohistochemistry; Keratins; Mouth Neoplasms

Sarcomatoid carcinoma of the upper aerodigestive tract continues to be one of the most difficult diagnostic challenges for surgical pathologists. Histogenesis has been settled in favor of a divergent (mesenchymal) differentiation of a carcinoma, most often a squamous cell carcinoma. Finding the carcinoma and/or its immunohistochemical marker in the metaplastic cells establishes the diagnosis. There are, however, lesions that can simulate sarcomatoid carcinomas to varying degrees, and in which neither a definable carcinoma nor immunohistochemical evidence of one can be found. Such lesions fall into several categories: 1. benign reactive lesions, 2. inflammatory myofibroblastic tumors, 3. sarcomas, usually low-grade, 4. atypical pseudosarcomatous proliferation. The clinicopathologic considerations of sarcomatoid carcinomas are presented in this context and include immunohistochemical findings, prognostic factors, and biologic course.

Topics: Carcinosarcoma; Cell Nucleus; Diagnosis, Differential; Digestive System Neoplasms; Epithelium; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Mesoderm; Mouth Neoplasms; Neoplasms, Muscle Tissue; Phenotype; Prognosis; Respiratory Tract Neoplasms

Merkel cell carcinoma (Mcc) is an uncommon and aggressive tumour with neuroendocrine features that occur predominantly in the head and neck region. The rarity of this tumour, especially when it arises in the oral mucosa, makes both early identification and standardisation of treatment difficult, particularly as regards complementary treatment. The availability of monoclonal antibodies with restricted specificity for some antigens thought to be related to neuroendocrine carcinomas, such as Merkel cell carcinoma, and ultrastructural studies offer some new leads to investigation. This has allowed, a greater number of these tumours to be discovered, thereby increasing the chances of effective management. A case of Mcc of the floor of the mouth is reported, together with the results of cytokeratin, neuron specific enolase and chromogranin immunohistochemistry.

Topics: Antibodies, Monoclonal; Carcinoma, Merkel Cell; Chromogranins; Fatal Outcome; Humans; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Mouth Floor; Mouth Neoplasms; Neoplasm Proteins; Phosphopyruvate Hydratase

In Sweden, snuff (locally known as snus), was introduced since the year 1637. Presently, Sweden has the highest per capita consumption and sale figures of snuff in the world, and the habit is becoming increasingly popular. Snus is manufactured into a dry form used in the nasal cavity and a moist form used in the oral cavity. Snus manufactured for oral use is a moist ground tobacco of Dark Kentucky or Virginia species mixed with an aqueous solution of water and other blending ingredients. This form of snuff is found in two types: (1) loose and (2) portion-bag-packed. These are the most widely used. The loose moist form (1-2 g a quid) is the most popular type consumed by 73% of the males, followed by the portion-bag-packed form (0.5-1 g a quid), consumed by 13% of the males, while 14% of the males are mixed users. The majority of snus users place the quid in the vestibular area of the upper lip, and the prevalence among persons 15 years of age or older in 15.9% among males and 0.2% among females. The pH of snus has declined from a previous range of 8-9 to a range of 7.8-8.5, moisture content ranges 35-60% and nicotine content is in the order of 5-11 mg/g dry wt tobacco-specific N-nitrosamines (TSNAs) in micrograms (N'-nitrosonornicotine: NNN 5-9; 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone: NNK 1-2; N'-nitrosoanatabine: NAT 2-5). In the Sudan, snuff, locally known as toombak, was introduced approximately 400 years ago. It is always processed into a loose moist form, and its use is widespread in the country. Tobacco used for manufacture of toombak is of the species Nicotiana rustica, and the fermented ground powder is mixed with an aqueous solution of sodium bicarbonate. The resultant product is moist, with a strong aroma, highly addictive and its use is widespread particularly among males. Its pH range is 8-11, moisture content ranges 6-60% and nicotine content is from 8 to 102 mg/g dry wt, and TSNAs contents in micrograms (NNN 420-1 550; NNK 620-7 870; NAT 20-290). Snus and toombak dippers develop a clinically and histologically characteristic lesion at the site of dipping. Probably due to control of the TSNAs in snus, this type of snuff is associated with a lower risk of cancer of the oral cavity (relative risk: RR 5-6-fold), whereas the risk for cancer of the oral cavity among toombak users was high (RR 7.3-73.0-fold). In conclusion, the two snuff products significantly differ in many aspects. Most notable differences are tobacco species, fermentatio

Topics: Carcinogens; Female; Genes, p53; Humans; Keratins; Male; Mouth Neoplasms; Mutation; Nitrosamines; Papillomaviridae; Plants, Toxic; Sudan; Sweden; Tobacco, Smokeless

The use of oral exfoliative cytology in clinical practice declined due to the subjective nature of its interpretation and because there may be only a small number of abnormal cells identifiable in a smear. The more recent application of quantitative techniques, together with advances in immunocytochemistry, have refined the potential role of cytology, stimulating a reappraisal of its value in the diagnosis of oral cancer. This review considers the influence of the quantitative analysis of cytomorphology, DNA analysis and other tumour markers applied to oral exfoliative cytological samples. These studies indicate that oral cytology may provide an important adjunct in the assessment of the patient with a potentially cancerous oral lesion.

Topics: Biomarkers, Tumor; Cell Transformation, Neoplastic; Cytodiagnosis; DNA, Neoplasm; Flow Cytometry; Humans; Keratins; Mouth Mucosa; Mouth Neoplasms; Pathology, Oral; Process Assessment, Health Care

A major limiting factor in the successful implementation of chemoprevention for oral cancer has been the lack of suitable endpoints to measure efficacy and success of clinical trials. To depend on the measure of the ultimate goal of such trials, namely cancer incidence as an endpoint has serious feasibility problems including a need for large numbers of participants, long periods of follow up and high costs. The application of selected biological markers as intermediate endpoints to reveal responses to chemopreventive regimens within a limited time frame is therefore an attractive concept. By serving as surrogates for cancer they could become valuable tools for monitoring patient compliance, defining the process of carcinogenesis and evaluating effects of the chemopreventive agents on tumour progression. This paper examines various biological markers of oral carcinogenesis and their relevance to chemoprevention trials. If validated, these biological markers could take oral cancer chemoprevention to new frontiers and help fulfil the ultimate goal of disease prevention through intervention.

Topics: Biomarkers, Tumor; Carotenoids; Chemoprevention; Clinical Trials as Topic; DNA, Neoplasm; Growth Substances; Humans; Keratins; Leukoplakia, Oral; Micronucleus Tests; Mouth Neoplasms; Outcome Assessment, Health Care; Retinoids

Six cases of squamous cell carcinoma arising in the head and neck of patients infected with the human immunodeficiency virus are described. This article reports the first two cases of primary intraosseous squamous cell carcinoma associated with infection with human immunodeficiency virus. Clinical presentation, results of imaging studies, histologic characteristics, therapies applied, and the clinical follow-up are described in detail for each of the six cases. These data are evaluated through a review of the current literature.

Topics: Acquired Immunodeficiency Syndrome; Adult; Biomarkers, Tumor; Carcinoma, Squamous Cell; CD4-CD8 Ratio; Female; Head and Neck Neoplasms; HIV Infections; Humans; Immunocompromised Host; Keratins; Male; Mandibular Neoplasms; Middle Aged; Mouth Neoplasms

The major value of intermediate filaments (IFs) in biological and applied research lies in their high order of cell and tissue specificity. This is particularly well illustrated in keratin (K) expression in various oral epithelia. Although the original class of IF is usually conserved in tissues after neoplastic transformation, epithelia show a tendency to shift their pattern of keratin expression in a manner which, while not predictable with precision, may sometimes be of diagnostic or prognostic significance. This review compares the keratins in normal oral epithelia, which show a mainly site-dependent expression, with those in squamous cell carcinoma. Key changes in the latter are the presence of simple epithelial keratins, K8 and K18 (occasional K7), reduced expression of differentiation-linked keratins (K1, K10, K4 and K13) and a tendency for down-regulation of primary keratins, K5 and K14. Moderate and severe dysplasias also tend to exhibit K8 and K18 with concomitant disordered expression of differentiation-linked keratins. There are reports of similar changes after neoplastic transformation in other mucosal sites and skin. Before this information can be applied diagnostically in immunocytochemical studies, the anti-keratin antibodies must be fully characterised and their interaction with the relevant tissue, both frozen and conventionally processed, should be evaluated.

Topics: Carcinoma, Squamous Cell; Humans; Intermediate Filaments; Keratins; Mouth Mucosa; Mouth Neoplasms; Neoplasm Proteins; Precancerous Conditions

A sensitive in situ hybridization test under low stringency conditions (LCS) with a set of digoxigenin-labeled human papillomavirus mixed probes (D-L HPV MP) revealed a positive reaction in 8 of 10 cases of oral verruca vulgaris (OVV). Ages ranged from 5 to 37 years with a mean of 14.5 years. 50% of all cases were located intraorally on the hard palate, followed in frequency by the commissures. These preliminary findings provide evidence of the role of HPV in OVV from a sample of the Venezuelan population. We show that in situ hybridization conducted under LSC is useful in HPV detection (regardless of the type) and the digoxigenin-labeling system is a rapid, relatively easy and specific method. In addition, this technique permits the retrospective evaluation of routinely processed material, thus widening the investigative spectrum for HPV.

Topics: Adolescent; Adult; Cell Nucleus; Child; Child, Preschool; Condylomata Acuminata; DNA Probes, HPV; Epithelium; Female; Humans; Keratins; Male; Mouth Diseases; Mouth Neoplasms; Nucleic Acid Hybridization; Papilloma; Papillomaviridae; Venezuela; Warts

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Benzoyl Peroxide; Cheek; Cocarcinogenesis; Cricetinae; Hyperplasia; Keratins; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Tetradecanoylphorbol Acetate

Human head and neck squamous cell carcinogenesis (SCC) is a common malignancy that appears to be related to continuous exposure to putative carcinogens or promoters such as tobacco and alcohol. To understand the mechanisms of the development of head and neck cancer and to test the efficiency of new therapeutic approaches, the characterization of an animal model system is necessary. The check-pouch carcinogenesis model in Syrian golden hamsters is probably the best known animal system that is most closely comparable with the development of premalignant and malignant lesions in human oral cancer. Furthermore, it is one of the most well-characterized animal system models for SCC. Our first approach to understanding the cellular and molecular changes that occur in the hamster cheek-pouch carcinogenesis process was to compare this model to the mouse-skin system, in which a number of critical events have been well characterized. We examined the sequential expression of hyperplasia, micronucleated cells, ornithine decarboxylase activity, polyamine levels, transglutaminase I activity, epidermal growth factor receptor levels, expression of several keratins, gamma-glutamyl transpeptidase, and nucleolar organizer regions. We suggest that these markers can be used to understand mechanisms of carcinogenesis and, in addition, can serve as alternative shorter end points in studies of chemoprevention. We also present preliminary molecular studies in the experimental oral model. We obtained a partial sequence of exon 2 of the Ha-ras gene and detected an A182----T transversion in codon 61 in hamster cheek-pouch SCC induced by 7,12-dimethylbenz(a)anthracene.

Topics: Animals; Biomarkers, Tumor; Cricetinae; Disease Models, Animal; gamma-Glutamyltransferase; Genes, Tumor Suppressor; Keratins; Mesocricetus; Mouth Neoplasms; Proto-Oncogenes

Topics: Epithelium; Fluorescent Antibody Technique; Humans; Keratins; Microscopy, Fluorescence; Mouth Mucosa; Mouth Neoplasms

Topics: Adolescent; Adult; Age Factors; Aged; Candidiasis, Oral; Child; Female; Follow-Up Studies; Humans; Hyperplasia; Keratins; Keratosis; Leukoplakia, Oral; Male; Middle Aged; Mouth Neoplasms; Nicotine; Precancerous Conditions; Retrospective Studies; Sex Factors; Smoking; Stomatitis

This article describes the use of tissue-engineered mucosal grafts instead of split-thickness skin grafts after freeing of the tongue in patients who had previous resection of an oral squamous cell carcinoma and initial primary wound closure.. Tissue-engineered mucosal grafts, up to 75 cm2 in size, were cultured from biopsy specimens of the hard palate in 6 patients, starting 3 to 4 weeks before the operation. After freeing of the tongue, the engineered mucosa was implanted on the wound surface by using vaseline gauze as carrier and fixed with an intraoral gauze wound dressing.. A good glossoalveolar sulcus was formed in 5 patients, resulting in good mobility of the tongue and a satisfactory denture-bearing surface. In 1 patient, there was a disturbance of wound healing, leading to severe shrinkage of the glossoalveolar sulcus and very limited improvement in tongue mobility. Preoperative bromodeoxyuridine (BrdU) labeling of the graft and postoperative immunohistochemical staining of biopsy specimens from the grafted areas with anti-BrdU showed that the cultured cells are integrated into the newly formed mucosal epithelium. Postoperative histologic investigations showed a differentiation process in the grafted mucosal epithelium, with a change in the expression of cytokeratins. At 6 months postoperatively, the typical pattern of normal nongrafted mucosa was regained.. This investigation provides evidence that tissue-engineered mucosal cells can serve as a graft for large intraoral wounds. Complete intraoral lining is quickly reestablished, and normal epithelial differentiation is seen in the graft area within a 6-month postoperative period.

Topics: Adult; Aged; Biomedical Engineering; Biopsy; Bromodeoxyuridine; Carcinoma, Squamous Cell; Cell Culture Techniques; Female; Histocytochemistry; Humans; Keratinocytes; Keratins; Male; Middle Aged; Mouth Floor; Mouth Mucosa; Mouth Neoplasms; Pilot Projects; Tongue; Vestibuloplasty

Oral squamous cell carcinoma (OSCC) is one of the most common malignancies affecting the head-and-neck region, regional lymph nodes being an important prognostication factor dictating the survival rate. Despite an array of modalities used, clinically, radiographically and routine histopathologically, the detection of micro-metastasis (2-3 mm tumour cell deposits) in the lymph nodes often escapes identification. The presence of few of these tumour epithelial cells in the lymph nodes drastically increases mortality and alters treatment plan. Hence, the identification of these cells is of major prognostic significance for a patient. Thus, the present study was aimed to evaluate and detect the efficacy of the immunohistochemical (IHC) marker [cytokeratin (CK) AE1/AE3] over routine Hematoxylin & eosin (H & E) staining in detecting micro-metastasis in the lymph nodes of OSCC cases.. The IHC marker CK cocktail (AE1/AE3) did not demonstrate any positive reactivity for the target antigen in all the 100 H & E stained lymph node sections evaluated in the present study.. This study was undertaken to check the efficacy of IHC (CK cocktail AE1/AE3) in the detection of micro-metastasis in lymph nodes that are found to be negative in routine H&E stained sections. The findings of this study suggest that the IHC marker AE1/AE3 did not prove to be useful to detect micro-metastasis in this study population.

Topics: Carcinoma, Squamous Cell; Eosine Yellowish-(YS); Head and Neck Neoplasms; Hematoxylin; Humans; Immunohistochemistry; Keratins; Lymph Nodes; Lymphatic Metastasis; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

Perineural invasion (PNI), a common occurrence in oral squamous cell carcinomas, is associated with poor survival. Consequently, these tumors are treated aggressively. However, diagnostic criteria of PNI vary and its role as an independent predictor of prognosis has not been established. To address these knowledge gaps, we investigated spatial and transcriptomic profiles of PNI-positive and PNI-negative nerves.. Tissue sections from 142 patients were stained with S100 and cytokeratin antibodies. Nerves were identified in two distinct areas: tumor bulk and margin. Nerve diameter and nerve-to-tumor distance were assessed; survival analyses were performed. Spatial transcriptomic analysis of nerves at varying distances from tumor was performed with NanoString GeoMx Digital Spatial Profiler Transcriptomic Atlas.. PNI is an independent predictor of poor prognosis among patients with metastasis-free lymph nodes. Patients with close nerve-tumor distance have poor outcomes even if diagnosed as PNI negative using current criteria. Patients with large nerve(s) in the tumor bulk survive poorly, suggesting that even PNI-negative nerves facilitate tumor progression. Diagnostic criteria were supported by spatial transcriptomic analyses of >18,000 genes; nerves in proximity to cancer exhibit stress and growth response changes that diminish with increasing nerve-tumor distance. These findings were validated in vitro and in human tissue.. This is the first study in human cancer with high-throughput gene expression analysis in nerves with striking correlations between transcriptomic profile and clinical outcomes. Our work illuminates nerve-cancer interactions suggesting that cancer-induced injury modulates neuritogenesis, and supports reclassification of PNI based on nerve-tumor distance rather than current subjective criteria.

Topics: Head and Neck Neoplasms; Humans; Keratins; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Staging; Peripheral Nerves; Prognosis; Retrospective Studies; Transcriptome

Transforming growth factor β (TGF-β) increases epithelial cancer cell migration and metastasis by inducing epithelial-mesenchymal transition (EMT). TGF-β also inhibits cell proliferation by inducing G1 phase cell-cycle arrest. However, the correlation between these tumor-promoting and -suppressing effects remains unclear. Here, we show that TGF-β confers higher motility and metastatic ability to oral cancer cells in G1 phase. Mechanistically, keratin-associated protein 2-3 (KRTAP2-3) is a regulator of these dual effects of TGF-β, and its expression is correlated with tumor progression in patients with head and neck cancer and migratory and metastatic potentials of oral cancer cells. Furthermore, single-cell RNA sequencing reveals that TGF-β generates two populations of mesenchymal cancer cells with differential cell-cycle status through two distinctive EMT pathways mediated by Slug/HMGA2 and KRTAP2-3. Thus, TGF-β-induced KRTAP2-3 orchestrates cancer cell proliferation and migration by inducing EMT, suggesting motile cancer cells arrested in G1 phase as a target to suppress metastasis.

Topics: Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; G1 Phase Cell Cycle Checkpoints; Gene Expression Regulation, Neoplastic; Humans; Keratins; Mouth Neoplasms; Transforming Growth Factor beta; Transforming Growth Factor beta1

Fanconi anemia (FA) patients frequently develop oral squamous cell carcinoma (OSCC). This cancer in FA patients is diagnosed within the first 3-4 decades of life, very often preceded by lesions that suffer a malignant transformation. In addition, they respond poorly to current treatments due to toxicity or multiple recurrences. Translational research on new chemopreventive agents and therapeutic strategies has been unsuccessful partly due to scarcity of disease models or failure to fully reproduce the disease. Here we report that Fanca gene knockout mice (Fanca

Topics: Animals; Carcinoma, Squamous Cell; Disease Models, Animal; Fanconi Anemia; Head and Neck Neoplasms; Keratins; Mice; Mice, Knockout; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

This study aims to perform differential protein expression analysis of serum samples from Oral Squamous Cell Carcinoma (OSCC) patients and healthy controls in search of potential diagnostic and/or prognostic biomarker(s).. OSCC is usually diagnosed late, which results in poor survival and high mortality. Identification of non-invasive prognostic biomarkers is of utmost importance for early diagnosis and proper management of the disease; hence we used a proteomic approach to identify potential biomarkers from serum.. Serum samples (OSCC n=45 and control n=30) were depleted, and proteins were separated using 2-D gel electrophoresis followed by identification by mass spectrometric analysis. Gene expression analysis of identified proteins in malignant and normal tissue was also performed to complement proteomics studies.. Among differentially expressed proteins, up-regulation of heat shock protein alpha (HSP90α) from the serum of oral cancer patients was observed. We also observed elevated levels of Haptoglobin (HP) along with downregulation of Type II keratin cytoskeletal 1(KRT1) and serum albumin (ALB) in oral cancer patients. Gene expression studies on identified proteins in malignant and normal tissue revealed a similar pattern with the exception of KRT1. We believe that elevated levels of serum HSP90 alpha might be used as a potential biomarker.. Our findings suggest a contribution of HSP90 alpha and other identified proteins in oral pathology as pro/anti-apoptotic modulators, thus considering their potential as predictive biomarkers.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Early Detection of Cancer; Gene Expression Regulation, Neoplastic; Haptoglobins; HSP90 Heat-Shock Proteins; Humans; Keratins; Mouth Neoplasms; Prospective Studies; Proteomics; Serum Albumin; Tandem Mass Spectrometry

Topics: Biomarkers, Tumor; Biopsy; Calcinosis; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Keratins; Mouth Neoplasms

Tumor budding (TB) is a promising prognostic marker in many cancers including oral squamous cell carcinoma. The evaluation of TB in preoperative diagnostic biopsies has been proven be possible; therefore, the association of TB with other morphological features can represent an important aid in the previous treatment decision. This study aims to evaluate TB in oral squamous cell carcinoma (OSCC) biopsies, assessing its association with other morphological characteristics of the sample. A total of 56 cases of OSCC were investigated. In hematoxylin and eosin-stained slides, morphological features including histopathological grading and mode of invasion were evaluated in the deep invasive front. Moreover, immunohistochemistry was performed with anti-multi-cytokeratin antibody helping in the identification of TB, which was graded as low-intensity or no TB and high-intensity TB. Descriptive and bivariate analyses were performed, and the level of significance was set at 5%. The tongue was the most-affected site with 29 (51.7 %) tumors. The predominant mode of invasion (27-48.2 %) was by groups of neoplastic cells without clear boundaries. Of the cases investigated, 37 (66.1 %) were high-intensity TB, which was associated with the mode of invasion of the tumors (p < 0.05). All cases with the worst mode of invasion showed high-intensity TB. Preliminary results showed the potential of morphological features, such as TB and mode of invasion, evaluated in diagnostic specimens of OSCC, aiding in the treatment decision to select patients who could benefit from more-aggressive treatments.

Topics: Adult; Biopsy; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

Bone morphogenetic protein 6 (BMP6) has unique properties regarding structure and function in supporting bone formation during development and adult life. Despite its known role in various malignant tumors, the prognostic significance of BMP6 expression in oral squamous cell carcinoma (OSCC) remains unknown. The aim of the study was to investigate immunohistochemical expression of BMP6 in OSCC in correlation with clinical and pathological parameters, disease recurrence and survival. In addition, we investigated other parameters in order to identify prognosticators of neck metastases and final outcome. The study included 120 patients with clinically T1-3N0 OSCC who were primarily surgically treated between 2003 and 2008. There were 99 (82.5%) male and 21 (17.5%) female patients. The five-year disease-specific survival for the whole cohort was 79.7%. Tumors smaller than 2 cm in diameter showed higher incidence of strong BMP6 expression. No statistical correlation was observed between other clinico-pathological factors and BMP6 expression. Expression of BMP6 was not associated with disease recurrence and survival. BMP6 may not serve as prognosticator of final outcome or recurrence in clinically node-negative OSCC subjects. In multivariate analysis predictors of poorer survival were positive surgical margin, moderate tumor cell differentiation and pathological involvement of levels IV and/or V.

Topics: Adult; Aged; Aged, 80 and over; Bone Morphogenetic Protein 6; Carcinoma, Squamous Cell; Cohort Studies; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Multivariate Analysis; Neoplasm Metastasis; Neoplasm Recurrence, Local; Prognosis

To identify cytokeratins (CK) of significant correlations with clinical and histopathologic prognostic parameters in oral and oropharyngeal squamous cell carcinoma (SCC).. The sample consisted of 100 cases retrieved from the archives of the Pathology Department/ King Hussein Cancer Center/Amman/ Jordan. Recorded data included: age, gender, location, grade, depth of invasion, the presence of epithelial dysplasia, tumor size, lymph node metastasis, number of positive lymph nodes, distant metastases, clinical stage, local recurrence, treatment modalities and 5-year survival rate. Immunohistochemical staining of 7 cytokeratins: 8, 10, 13, 14, 16, 18, and 19 was performed using standard protocols. Stained sections were digitized and analyzed using ImageJ-color deconvolution to identify the percentage of cytokeratin-positive area (score). Statistical tests used were: student t-test, analysis of variance, bivariate analysis and logistic regression.. Lower CK8,18, 19 scores correlated with lower 5-year survival rate. Higher CK19 and lower CK 10, 14, 16 scores were associated with distant metastasis. Increased CK8, 18, 19 scores correlated with higher stage and with higher depth of invasion. Increased CK18 scores correlated with increased local recurrence. Higher CK10, 13, 16 scores correlated with well-differentiated grade. Higher CK19 and lower CK16 scores were associated with adjacent epithelial dysplasia. Regression analysis showed that better 5-year survival rate was significantly correlated with increased CK16, decreased CK18 and 19 scores.. Expression scores of a panel of cytokeratin are potential prognostic indicators for 5-year survival and correlates with other prognostic parameters.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Carcinoma, Squamous Cell; Female; Gingiva; Humans; Immunohistochemistry; Jordan; Keratin-10; Keratin-13; Keratin-14; Keratin-16; Keratin-18; Keratin-19; Keratin-8; Keratins; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Oropharynx; Prognosis; Regression Analysis; Survival Rate; Young Adult

Keratinization is a routinely reported histologic feature in head and neck cancer. In contrast to numerous clinicopathologic parameters, the prognostic value of keratinization in oral squamous cell carcinoma (OSCC) is rarely reported in the literature. The purpose of this study was to review the outcome of patients with OSCC with a special focus on the degree of keratinization.. In this retrospective cohort study, we evaluated the medical records at the Department of Oral and Maxillofacial Surgery, Jena University Hospital, and investigated the outcome of patients with OSCC with disease-free survival and disease-specific survival according to the degree of keratinization. This research also analyzed common clinical and histologic parameters such as age, gender, tumor site, T category, N category, resection margin, lymphovascular invasion, and extracapsular spread. Descriptive statistics were performed, and survival was calculated by the Kaplan-Meier method. Prognostic factors were analyzed by multivariate Cox analysis.. In the sample of 151 OSCC patients, with a median age of 57.5 years and a male-female ratio of 4.03:1, 119 had tumors with no or low keratinization (K0 to K2) and 32 had tumors with good or high keratinization (K3 or K4). More recurrences were seen in patients with OSCC with low keratinization (P = .0008). The 5-year disease-free survival rate was significantly decreased for OSCC with low keratinization (52.9%) compared with good or high keratinization (93.2%) (P = .0008). The 5-year disease-specific survival rate was reduced to 66.1% (P = .0136) for patients with OSCC with low keratinization. Multivariate analysis showed that extracapsular spread (P = .001) and keratinization (P = .002) are independent, significant prognostic factors for recurrence in OSCC.. Besides extracapsular spread, the degree of keratinization seems to be an important prognostic factor for recurrence and survival in OSCC. Our results indicate that the degree of keratinization should be considered in decisions regarding treatment and prognosis for OSCC.

Topics: Carcinoma, Squamous Cell; Female; Humans; Keratins; Lymphatic Metastasis; Male; Margins of Excision; Middle Aged; Mouth Neoplasms; Neoplasm Grading; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Retrospective Studies; Survival Rate

Tumor budding is a morphological marker of cancer invasion, defined as the presence of isolated or small clusters of neoplastic cells at the tumor invasive front. This study aimed to evaluate the association between intensity of tumor budding and cell proliferation in oral squamous cell carcinoma (OSCC).. Immunohistochemistry was employed in 163 OSCC samples to detect the cell proliferation marker Ki-67 and multicytokeratin (to identify OSCC cells in tumor budding evaluation). The Mann-Whitney test was used to evaluate differences in the cell proliferation index between samples with high-intensity tumor budding and samples with low-intensity or no tumor budding. In samples with high-intensity tumor budding, the Wilcoxon test was used to evaluate differences in the cell proliferation index between the budding area and the area outside the budding. The chi-square test assessed the association between cell proliferation index and intensity of tumor budding.. The cell proliferation index was higher in samples with high-intensity tumor budding than in samples with low-intensity or no tumor budding (P < .05). Tumors with high-intensity tumor budding showed a higher cell proliferation index in the budding area than in the area outside the budding (P < .05). Finally, samples showing high-intensity tumor budding were associated with high cell proliferation index (P < .05).. Cell proliferation is positively associated with intensity of tumor budding in OSCC. Moreover, in tumors showing high-intensity tumor budding, the budding area is the location of higher cell proliferation. These findings reinforce the hypothesis that tumor budding is associated with the biological behavior of OSCC.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Proliferation; Female; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Male; Middle Aged; Mouth Neoplasms; Pathology, Clinical; Staining and Labeling

Keratin 76 (Krt76) is expressed in the differentiated epithelial layers of skin, oral cavity and squamous stomach. Krt76 downregulation in human oral squamous cell carcinomas (OSCC) correlates with poor prognosis. We show that genetic ablation of Krt76 in mice leads to spleen and lymph node enlargement, an increase in regulatory T cells (Tregs) and high levels of pro-inflammatory cytokines. Krt76

Topics: 5'-Nucleotidase; Animals; Antigens, CD; Apyrase; Cell Line, Tumor; Female; Flow Cytometry; Fluorescent Antibody Technique; Humans; In Situ Hybridization, Fluorescence; In Vitro Techniques; Keratins; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Mutant Strains; Mouth Neoplasms; Stomach Neoplasms; T-Lymphocytes, Regulatory

Multipotent stromal cells (MSCs) are envisioned as a powerful therapeutic tool. As they home into tumors, secrete trophic and vasculogenic factors, and suppress immune response their role in carcinogenesis is a matter of controversy. Worldwide oral squamous cell carcinoma (OSCC) is the fifth most common epithelial cancer. Our aim was to determine whether MSC administration at precancerous stage modifies the natural progression of OSCC. OSCC was induced in Syrian hamsters by topical application of DMBA in the buccal pouch. At papilloma stage, the vehicle or 3×10

Topics: Animals; Apoptosis; Bone Marrow Cells; Carcinoma, Squamous Cell; Caspase 3; Cell Dedifferentiation; Cell Line, Tumor; Cell Proliferation; Cricetinae; Disease Progression; Down-Regulation; Epithelial Cells; Female; Hyperplasia; Immunophenotyping; Keratins; Ki-67 Antigen; Leukocyte Common Antigens; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mesocricetus; Mouth Neoplasms; Papilloma; Phenotype; Transcriptome; Transplantation, Homologous

The cytology of oral squamous cell carcinoma (SCC) is challenging because oral SCC cells tend to be well differentiated and lack nuclear atypia, often resulting in a false negative diagnosis. The purpose of this study was to establish practical cytological parameters specific to oral SCCs.. We reviewed 123 cases of malignancy and 53 of non-neoplastic lesions of the oral mucosa, which had been diagnosed using both cytology and histopathology specimens. From those, we selected 12 SCC and 4 CIS cases that had initially been categorized as NILM to ASC-H with the Bethesda system, as well as 4 non-neoplastic samples categorized as LSIL or ASC-H as controls, and compared their characteristic findings. After careful examinations, we highlighted five cytological parameters, as described in Results. Those 20 cytology samples were then reevaluated by 4 independent examiners using the Bethesda system as well as the 5 parameters.. Five cytological features, (i) concentric arrangement of orangeophilic cells (indicating keratin pearls), (ii) large number of orangeophilic cells, (iii) bizarre-shaped orangeophilic cells without nuclear atypia, (iv) keratoglobules, and (v) uneven filamentous cytoplasm, were found to be significant parameters. All malignant cases contained at least one of those parameters, while none were observed in the four non-neoplastic cases with nuclear atypia. In reevaluations, the Bethesda system did not help the screeners distinguish oral SCCs from non-neoplastic lesions, while use of the five parameters enabled them to make a diagnosis of SCC.. Recognition of the present five parameters is useful for oral SCC cytology. Diagn. Cytopathol. 2017;45:406-417. © 2017 Wiley Periodicals, Inc.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Nucleus; Cytoplasm; Diagnosis, Differential; Female; Histocytochemistry; Humans; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms

We have developed an automatic and objective method for detecting human oral squamous cell carcinoma (OSCC) tissues with Raman microspectroscopy. We measure 196 independent Raman spectra from 196 different points of one oral tissue sample and globally analyze these spectra using a Multivariate Curve Resolution (MCR) analysis. Discrimination of OSCC tissues is automatically and objectively made by spectral matching comparison of the MCR decomposed Raman spectra and the standard Raman spectrum of keratin, a well-established molecular marker of OSCC. We use a total of 24 tissue samples, 10 OSCC and 10 normal tissues from the same 10 patients, 3 OSCC and 1 normal tissues from different patients. Following the newly developed protocol presented here, we have been able to detect OSCC tissues with 77 to 92% sensitivity (depending on how to define positivity) and 100% specificity. The present approach lends itself to a reliable clinical diagnosis of OSCC substantiated by the "molecular fingerprint" of keratin.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Humans; Keratins; Mouth Neoplasms; Spectrum Analysis, Raman

Granular cell tumor (GCT) of the oral cavity is a benign lesion. Half of oral GCTs demonstrate pseudocarcinomatous hyperplasia (PCH) of the mucosa which can mimic invasive islands of oral squamous cell carcinoma (SCC). Such similarity can be confusing when diagnosing or evaluating the two conditions, potentially leading to misdiagnosis or misclassification. Indeed, several misdiagnosed cases of oral GCT have been reported in the literature as OSCC or malignant oral GCT that resulted in unnecessary aggressive treatment for the affected patients. The aim of this study was to investigate if the cytokeratin pattern of the PCH can help in differentiating GCT from oral SCC. To distinguish between these two entities, we examined 12 patient specimens of oral GCT-PCH and oral SCC histologically and via immunohistochemistry (IHC) for CK13, CK17 and P75. The results suggest that the cytokeratin profile of PCH is similar to that of oral SCC. Therefore, consideration of IHC findings for epithelial markers alone may lead to erroneous diagnosis; thus, the presence of the granular tumor underneath the PCH and its immunopositivity for P75 or other neural definition markers can be essential to identify the underlying tumor and exclude oral SCC. Finally we recommend more studies on the molecular biology of PCH to understand how it can mimic oral SCC histologically without harboring its malignant phenotype clinically, which could have significant translational potential for understanding invasive oral SCC.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Diagnosis, Differential; Granular Cell Tumor; Humans; Hyperplasia; Immunoenzyme Techniques; Keratins; Mouth Neoplasms; Neoplasm Staging; Prognosis

Squamous cell carcinoma (SSC) of the head and neck is the sixth most common cancer and is rarely diagnosed in early stages. The transcription factor Krϋppel-like factor 4 (Klf4) suppresses cell proliferation and promotes differentiation. Inducible mice carrying an oral-specific ablation of Klf4 (K14-CreER(tam) /Klf4(flox/flox) ) develop mild dysplastic lesions and abnormal differentiation in the tongue. Aiming to analyze whether Klf4 cooperate in oral chemical carcinogenesis,we applied 4-nitroquinoline 1-oxide (4NQO), a tobacco surrogate, to this conditional Klf4 knockout mice.. K14-CreER(tam) /Klf4(flox/flox) and control mice were treated with 4NQO for 16 weeks and monitored until week 30. Histopathological samples were used for diagnostic purposes and immunofluorescence detection of epithelial differentiation markers.. 4NQO-treated K14-CreER(tam) /Klf4(flox/flox) mice (Klf4KO 4NQO) showed a significant weight loss and developed more severe dysplastic lesions than control mice with 4NQO (P < 0.005). The Klf4KO 4NQO showed a tendency to higher incidence of oral SCC and a marked keratinization pattern in dysplasias, in situ carcinomas and SCC. Also, tongues derived from Klf4KO 4NQO mice exhibited reduced terminal differentiation as judged by cytokeratin 1 staining when compared with 4NQO-treated controls.. Klf4 ablation results in more severe dysplastic lesions in oral mucosa, with a tendency to higher incidence of SCC, after chemical carcinogenesis. We show here, in a context similar to the human carcinogenesis, that absence of Klf4 accelerates carcinogenesis and correlates with the absence of cytokeratin 1 expression. These results suggest a potential role for KLF4 as a tumor suppressor gene for the tongue epithelium.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinogenesis; Carcinogens; Carcinoma, Squamous Cell; Cell Differentiation; Disease Models, Animal; Gene Expression; Head and Neck Neoplasms; Keratins; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Mice; Mice, Inbred C57BL; Mice, Knockout; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Squamous Cell Carcinoma of Head and Neck; Tongue Neoplasms

Oral squamous cell carcinomas (OSCC) induced in F344 rats by 4-nitroquinoline-1-oxide (4-NQO) demonstrate considerable phenotypic similarity to human oral cancers. Gene expression studies (microarray and PCR) were coupled with methylation analysis of selected genes to identify molecular markers of carcinogenesis in this model and potential biochemical and molecular targets for oral cancer chemoprevention. Microarray analysis of 11 pairs of OSCC and site-matched phenotypically normal oral tissues from 4-NQO-treated rats identified more than 3500 differentially expressed genes; 1735 genes were up-regulated in rat OSCC versus non-malignant tissues, while 1803 genes were down-regulated. In addition to several genes involved in normal digestion, genes demonstrating the largest fold increases in expression in 4-NQO-induced OSCC include three lipocalins (VEGP1, VEGP2, LCN2) and three chemokines (CCL, CXCL2, CXCL3); both classes are potentially druggable targets for oral cancer chemoprevention and/or therapy. Down-regulated genes in 4-NQO-induced OSCC include numerous keratins and keratin-associated proteins, suggesting that alterations in keratin expression profiles may provide a useful biomarker of oral cancer in F344 rats treated with 4-NQO. Confirming and extending our previous results, PTGS2 (cyclooxygenase-2) and several cyclooxygenase-related genes were significantly up-regulated in 4-NQO-induced oral cancers; up-regulation of PTGS2 was associated with promoter hypomethylation. Rat OSCC also demonstrated increased methylation of the first exon of APC2; the increased methylation was correlated with down-regulation of this tumor suppressor gene. Overexpression of pro-inflammatory chemokines, hypomethylation of PTGS2, and hypermethylation of APC2 may be causally linked to the etiology of oral cancer in this model.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinoma, Squamous Cell; Chemokines; Cyclooxygenase 1; Cyclooxygenase 2; DNA Methylation; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Keratins; Lipocalins; Male; Membrane Proteins; Mouth Neoplasms; Oligonucleotide Array Sequence Analysis; Promoter Regions, Genetic; Rats, Inbred F344; Tongue; Transcriptome

To evaluate the effectiveness of sentinel lymph node biopsy (SLNB) as an optimal staging method in oral and oropharyngeal squamous cell carcinoma (OOSCC) and the impact of the extent of SLN involvement on the decision for neck dissection (ND).. A prospective cohort study was performed in 96 consecutive patients with stage T1 to T4N0M0 OOSCC (mean follow-up, 62.9 months). SLN localization was determined using cervical lymphoscintigraphy and single-photon emission computed tomography. Patients underwent SLNB examination and ND. The ND specimen was investigated by hematoxylin and eosin (H&E) staining and the SLNs were investigated using H&E staining and step-serial sectioning and cytokeratin antibodies AE1 and AE3. The statistical study calculated the sensitivity and negative predictive value (NPV). The sample size of 96 patients was calculated for a 95% confidence interval with an accuracy of ±2% and an estimated a priori sensitivity of 99% compared with the benchmark. The impact of extent of SLN involvement on the decision for ND was analyzed by χ(2) test. A logistic regression model was used to assess the association of predictor variables with SLN involvement and neck disease.. The diagnostic accuracy, sensitivity, NPV, and negative likelihood ratio were 95%, 88%, 94%, and 0.06. The statistical comparison between the extent of metastatic involvement of the SLN and neck disease was important for SLN macrometastasis (odds ratio = 11.9), but not for SLN micrometastasis (odds ratio = 0.93).. SLNB examination is an excellent staging method in OOSCC. The present data indicate a very small risk of additional lymph node metastasis with SLN micrometastasis.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cohort Studies; Decision Making; Female; Follow-Up Studies; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Lymphoscintigraphy; Male; Middle Aged; Mouth Neoplasms; Neck Dissection; Neoplasm Staging; Oropharyngeal Neoplasms; Predictive Value of Tests; Prospective Studies; Radiopharmaceuticals; Sensitivity and Specificity; Sentinel Lymph Node Biopsy; Technetium; Tomography, Emission-Computed, Single-Photon

The invasion of cancer cells into the surrounding normal tissue is one of the defining features of cancer. While the phenomena of tumour budding, epithelial-mesenchymal transition and the presence of myofibroblasts have independently been shown to be related to a poor prognosis of oral carcinomas, their relationship has not been examined in detail.. Paraffin-embedded tissues from 28 patients with oral squamous cell carcinomas were stained with antibodies to cytokeratin, α-SMA, vimentin, E-cadherin, N-cadherin and Twist and evaluated for their expression in relation to invasive cancer cells and the surrounding tumour stroma.. A direct, histological relationship between invading, budding tumour cells and myofibroblasts was occasionally seen but was not a general feature. Most of the budding tumour cells at the invasive front had a decreased expression of E-cadherin, but we did not find that this was associated with a consistent or clear increase in either N-cadherin or vimentin. We therefore suggest that the budding of tumour cells is not dependent upon either myofibroblasts or a complete epithelial-mesenchymal transition and that these phenomena most likely represent separate processes in tumour progression.

Topics: Actins; Animals; Cadherins; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; Goats; Head and Neck Neoplasms; Humans; Keratins; Mice; Mouth Neoplasms; Muscle, Smooth; Myofibroblasts; Paraffin Embedding; Prognosis; Rabbits; Single-Cell Analysis; Squamous Cell Carcinoma of Head and Neck; Twist-Related Protein 1; Vimentin

Cytokeratins (CK) are abundant in keratinized cells, particularly CK14 and CK19, which are expressed in stratified squamous epithelial cells. In this study, expression of CK14 and 19 was examined in human epithelial and dysplastic tissues. Surgical specimens from patients with clinically diagnosed oral leukoplakia or early cancer were stained with hematoxylin and eosin and classified into normal, low grade dysplasia (LGD), high grade dysplasia (HGD), or squamous cell carcinoma (SCC). The sections were examined by immunostaining and reverse transcription-polymerase chain reaction (RT-PCR) for CK14 and CK19. Expression and the results of RT-PCR for CK14 showed a decrease in the order of LGD, HGD, and SCC, whereas those of CK19 showed an increase in that order. These results suggest that decreased expression of CK14 and increased expression of CK19 serve as indicators of potential for malignant transformation.

Topics: Carcinogenesis; Carcinoma, Squamous Cell; Epithelial Cells; Humans; Keratin-14; Keratin-19; Keratins; Mouth Neoplasms

Oral squamous cell carcinoma (OSCC) has contributed 90% of oral cancer worldwide. In situ histological evaluation of tissue sections is the gold standard for oral cancer detection. Formation of keratinization and keratin pearl is one of the most important histological features for OSCC grading. This paper aims at developing a computer assisted quantitative microscopic methodology for automated identification of keratinization and keratin pearl area from in situ oral histological images. The proposed methodology includes colour space transform in YDbDr channel, enhancement of keratinized area in most significant bit (MSB) plane of Db component, segmentation of keratinized area using Chan-Vese model. The proposed methodology achieves 95.08% segmentation accuracy in comparison with (manually) experts-based ground truths. In addition, a grading index describing keratinization area is explored for grading OSCC cases (poorly, moderately and well differentiated).

Topics: Animals; Carcinoma, Squamous Cell; Diagnostic Imaging; Humans; Keratins; Mouth Neoplasms; Neoplasm Grading

Five-year survival rates for oral squamous cell carcinoma (OSCC) are 30% and the mortality rate is 50%. Immunohistochemistry panels are used to evaluate proliferation, vascularization, apoptosis, HPV infection, and keratin expression, which are important markers of malignant progression. Keratins are a family of intermediate filaments predominantly expressed in epithelial cells and have an essential role in mechanical support and cytoskeleton formation, which is essential for the structural integrity and stability of the cell. In this study, we analyzed the expressions of keratins 17 and 19 (K17 and K19) by immunohistochemistry in tumoral and non-tumoral tissues from patients with OSCC. The results show that expression of these keratins is higher in tumor tissues compared to non-tumor tissues. Positive K17 expression correlates with lymph node metastasis and multivariate analysis confirmed this relationship, revealing a 6-fold increase in lymph node metastasis when K17 is expressed. We observed a correlation between K17 expression with disease-free survival and disease-specific death in patients who received surgery and radiotherapy. Multivariate analysis revealed that low expression of K17 was an independent marker for early disease relapse and disease-specific death in patients treated with surgery and radiotherapy, with an approximately 4-fold increased risk when compared to high K17 expression. Our results suggest a potential role for K17 and K19 expression profiles as tumor prognostic markers in OSCC patients.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease-Free Survival; Epithelial Cells; Female; Humans; Keratins; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neoplasm Recurrence, Local; Prognosis

Oral squamous cell carcinoma (OSCC) is one of the most prevalent cancers worldwide. Laminin-5 gamma 2 chain (laminin-5 γ2) is a protein associated to a migratory phenotype in epithelial neoplastic cells. Stromal myofibroblasts also play a significant role in tumor invasion, due to its ability to modify the extracellular matrix. Tumor budding is a morphologic marker of tumor invasion. The aim of this study was to evaluate the expression of laminin-5 γ2 in OSCC and its association with intensity of tumor budding and density of stromal myofibroblasts.. Paraffin-embedded archival samples of 57 OSCC patients were evaluated. Immunohistochemistry was employed to detect laminin-5 γ2, alpha smooth muscle actin (marker of stromal myofibroblasts), and multicytokeratin (to identify OSCC cells in tumor budding evaluation). Laminin-5 γ2 expression and its association with intensity of tumor budding and density of stromal myofibroblasts were analyzed. Association among intensity of tumor budding and density of stromal myofibroblasts was also evaluated.. Higher laminin-5 γ2 expression was associated with high-intensity tumor budding (P < 0.05) and with higher density of stromal myofibroblasts (P < 0.05). Moreover, high-intensity tumor budding was associated with higher density of stromal myofibroblasts (P < 0.05).. In OSCC, higher laminin-5 γ2 expression is associated with high-intensity tumor budding and with higher density of stromal myofibroblasts, suggesting that this expression is related to the establishment of an invasive phenotype of neoplastic cells and a permissive environment for tumor invasion in this neoplasia.

Topics: Actins; Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Count; Female; Humans; Immunohistochemistry; Keratins; Laminin; Male; Middle Aged; Mouth Neoplasms; Myofibroblasts; Neoplasm Invasiveness

We have reported that neutrophilic infiltration was associated with round-shaped dyskeratosis foci, a kind of keratin pearl, of oral carcinoma in situ and that those inflammatory cells are recruited from intra-epithelially entrapped blood vessels. Based on these lines of evidence, we have formulated a hypothesis that keratin pearls are terminally degraded by neutrophils. To confirm this hypothesis, we investigated immunohistochemically stepwise degradation of keratin pearls in oral squamous cell carcinoma (SCC) to clarify any other type scavenger cells in addition to neutrophils are involved in this particular degradation process.. Neutrophils (neutrophil elastase) and macrophage subpopulations (CD68, CD163 and CD204) were immunohistochemically localized in 30 cases of oral SCC with typical round-shaped keratin pearls. SCC cells were revealed by immunohistochemistry for keratin (K) 17, and blood vessels were demonstrated by CD31.. Keratin pearl degradation process was divided into four steps: (i) intact stage: no macrophage infiltration but minimal neutrophils were found in keratin pearls; (ii) neutrophil recruit stage: no macrophage infiltration but focal neutrophilic infiltration within the pearls; (iii) neutrophil predominant stage: dense neutrophil infiltration with minimal macrophages and segregated keratinized cancer cells strongly positive for K17; and (iv) macrophage predominant stage: dense infiltration of CD68-, CD163 (mononuclear)- and CD204 (multinucleated)-positive macrophages engulfing detached keratinized SCC cells.. Keratin pearl degradation in oral SCC is strictly regulated by two types of scavenger cells: neutrophils, which perform initial tasks, and macrophages, which reciprocally take over from neutrophils the role to finalize the degradation processes.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Carcinoma, Squamous Cell; Humans; Keratin-17; Keratins; Leukocyte Elastase; Macrophage Activation; Macrophages; Mouth Neoplasms; Neutrophil Infiltration; Neutrophils; Platelet Endothelial Cell Adhesion Molecule-1; Proteolysis; Receptors, Cell Surface; Receptors, Scavenger; Scavenger Receptors, Class A

An adult, gravid, female pigtailed macaque (Macaca nemestrina) presented for facial swelling centered on the left mandible that was approximately 5 cm wide. Differential diagnoses included infectious, inflammatory, and neoplastic origins. Definitive antemortem diagnosis was not possible, and the macaque's condition worsened despite supportive care. Necropsy findings included a mandibular mass that was locally invasive and expansile, encompassing approximately 80% of the left mandibular bone. The mass replaced portions of the soft palate, hard palate, sinuses, ear canal, and the caudal-rostral calvarium and masseter muscle. Histologically, the mass was a neoplasm that was poorly circumscribed, unencapsulated, and infiltrative invading regional bone and soft tissue. The mass consisted of polygonal squamous epithelial cells with intercellular bridging that breached the epithelial basement membrane and formed invasive nests, cords, and trabeculae. The mitotic rate averaged 3 per 400× field of view, with occasional bizarre mitotic figures. Epithelial cells often exhibited dyskeratosis, and the nests often contained compact lamellated keratin (keratin pearls). The neoplasm was positive via immunohistochemistry for pancytokeratin, variably positive for S100, and negative for vimentin, smooth muscle actin, and desmin. The gross, histologic, and immunohistochemical findings were consistent with an aggressive oral squamous cell carcinoma. The neoplasm was negative via PCR for papilloma virus. In general, neoplasia in macaques is rare. Although squamous cell carcinomas are one of the most common oral neoplasia in many species, to our knowledge this case represents the first reported oral squamous cell carcinoma in a pigtailed macaque.

Topics: Animals; Carcinoma, Squamous Cell; Fatal Outcome; Female; Immunohistochemistry; Keratins; Macaca nemestrina; Monkey Diseases; Mouth Neoplasms

Oral squamous cell carcinoma (OSCC) and canine acanthomatous ameloblastoma (CAA) represent two epithelium-derived neoplasms that affect the oral cavity of dogs. The expression of cytokeratins (CKs) and calretinin has been previously established in the canine tooth bud and odontogenic tumours. The aim of this study was to characterize the CK and calretinin expression profile of OSCC in comparison to CAA and canine tooth bud tissues. Samples from 15 OSCC and 15 CAA cases, as well as 6 tooth buds and 2 normal gingival tissues were examined. OSCC CK expression was consistent with the CK expression profile of CAA and canine tooth bud tissue. Calretinin was positively expressed in 10 of 15 OSCC cases, with 5 cases demonstrating high staining intensity. Only 2 of 15 CAA cases demonstrated mild-moderate staining intensity. The statistically significant difference in staining pattern and intensity of calretinin in OSCC and CAA can help distinguish between these two tumour types.

Topics: Ameloblastoma; Animals; Antibodies, Monoclonal; Calbindin 2; California; Carcinoma, Squamous Cell; Dog Diseases; Dogs; Immunohistochemistry; Jaw Neoplasms; Keratins; Mouth Neoplasms; Tooth; Universities

The objective of this study was to evaluate the potential detection of circulating tumor cells (CTCs) using the CellSearch (CS) Assay™ in patients with locally advanced head and neck squamous cell carcinoma (HNSCC) and then to identify the clinical factors predictive of the presence of CTCs. The presence and number of CTCs were determined using the CS system before treatment, and in 10 healthy individuals (control group). The CS system was able to successfully identify the presence of CTCs in 8 of 49 patients (16 %) before therapy. No CTC was found in the control group. CTCs were detected before therapy in 1 of 19 patients (5 %) with N0 tumor and in 7 of 30 patients (23 %) with N1-2c tumor (p = 0.12; Fisher's exact test). CTCs were identified in a relatively low proportion of patients with locally advanced HNSCC.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Cell Adhesion Molecules; Cell Count; Epithelial Cell Adhesion Molecule; Female; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Neoplastic Cells, Circulating; Oropharyngeal Neoplasms; Prognosis; Squamous Cell Carcinoma of Head and Neck

The purpose of this study was to assess the predictability of sentinel lymph node biopsy (SNB) for oral squamous cell carcinoma (OSCC) when pathologic processing is performed without serial step sectioning.. We prospectively enrolled 36 patients with T1 or T2 cN0 OSCC into this institutional review board-approved prospective cohort study, and they underwent gamma probe-guided SNB in addition to selective neck dissection. The rate of patients with negative SNB results whose neck dissection was also negative for metastasis (negative predictive value) was the primary endpoint.. Of the 28 patients whose sentinel lymph nodes were found to be pathologically and clinically node negative by routine hematoxylin-eosin stain and immunohistochemistry, 27 were found to have no other pathologically positive nodes, corresponding to a negative predictive value of 96%.. The results of this study suggest that SNB performed without the use of thin serial step sectioning may accurately predict neck stage in OSCC.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cohort Studies; Coloring Agents; Eosine Yellowish-(YS); Fluorescent Dyes; Hematoxylin; Humans; Immunohistochemistry; Keratins; Lymph Nodes; Lymphatic Metastasis; Microtomy; Middle Aged; Mouth Neoplasms; Neck Dissection; Neoplasm Staging; Predictive Value of Tests; Prospective Studies; Radionuclide Imaging; Radiopharmaceuticals; Sentinel Lymph Node Biopsy; Technetium Tc 99m Sulfur Colloid; Young Adult

In humans, activation-induced cytidine deaminase (AID) expression results due to inflammation and this deaminase activity is also involved in carcinogenesis. The aim of this study is to investigate the correlation between AID expression and the clinical classification of oral cancer tissues.. The current study investigated the correlation between AID expression and the clinical classification of oral cancer tissues from 27 patients who underwent surgical resection using immunohistochemistry. Specific AID expression and its induction by cytokine stimulation were investigated in cultured HSC oral cancer cell lines by reverse transcriptase PCR.. AID expression was detected in 10 of 27 specimens (37.0%). AID expression was more frequently detected in early-stage cancer, especially in early stage T, than in late-stage cancer (T1/T2 vs. T3/4; P = 0.0493, N0 vs. N1/2/3; P = 0.0793). HSC-2, a nonmetastatic oral cancer cell line, abundantly expressed endogenous AID, whereas no such expression was observed in HSC-3, a metastatic oral cancer cell line. Moreover, AID expression was substantially induced in HSC-2 cells by stimulation of an inflammation-related cytokine, TNF-α.. Aberrant AID expression in the oral epithelium would contribute to the initiation of oral squamous cell carcinoma. Avoiding persistent AID inducible condition such as frequent cleaning of oral cavity would play an important role for the prevention of developing oral cancer.

Topics: Carcinoma, Squamous Cell; Cell Line, Tumor; Cytidine Deaminase; Humans; Keratins; Mouth Neoplasms; Neoplasm Staging; Tumor Necrosis Factor-alpha; Up-Regulation

Progression of oral carcinomas associates with aberrant activation and inactivation of molecules that work in established or unknown pathways. Although mucosa‑associated lymphoid tissue 1 (MALT1) expressed in normal oral epithelium is inactivated in the aggressive subset of carcinomas with worse prognosis, phenotypic changes of carcinoma cells upon the loss of expression is unknown. We performed a proteomic analysis to identify MALT1‑regulated proteins in oral carcinoma cells. Four different keratins were included in the ten most abundantly changed proteins. K8/18 were upregulated in MALT1 stably‑expressing carcinoma cells and K5/14 in MALT1‑marginal control cells. K8/18 upregulation and K5/14 downregulation were MALT1 dose‑dependent and observed in a series of oral carcinoma cells. MALT1 suppressed cell proliferation (0.52-fold, P<0.01) and its dominant-negative form stimulated it (1.33-fold, P<0.01). The decreased proliferation associated with reduction of cyclin D1, which was recovered by the short interfering RNA against MALT1. Taken together, loss of MALT1 expression alters keratin expression and enhances proliferation of carcinoma cells, and may progress oral carcinomas into the advanced state.

Topics: Carcinoma; Caspases; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Epithelium; Gene Expression Regulation, Neoplastic; Humans; Keratins; Mouth Neoplasms; Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein; Mucous Membrane; Neoplasm Proteins; Proteomics; Up-Regulation

To evaluate the expression of cytokeratin 19(CK19) and connexin 43(Cx43) in various stages of oral carcinogenesis and investigate the relationship of CK19 and Cx43 in the process of oral cancer.. 4-nitroquinoline-1-oxide(4NQO) was used to induce oral carcinogenesis in the mucosa of SD rats and immunohistoche-mical technique was used to study the expression of CK19 and Cx43 in various stages of oral carcinogenesis.. The CK19 positive staining were distributed in the basal cell layer in the normal rat lingual mucosa. While CK19 positive staining were distributed in cytoplasm of supra-basal layers in the mild dysplasia, moderate dysplasia and severe dysplasia. In oral squamous cell carcinoma(OSCC) tissue, CK19 were expressed in all the stratum of epithelium. The positive rate of CK19 in normal, mild, moderate, severe dysplasia and OSCC tissues were respectively 30.00%, 50.00%, 58.33%, 80.00%, and 91.67%. With the lesions getting worse, the positive rate and the intensity of CK19 raised significantly (P<0.05). In normal tongue mucosa, Cx43 proteins were mainly expressed in the membrane of the epithelial cells of the rat tongue. It was weakly positive in the basal cell layer, increased in the stratum spinosum and stratum granulosum, and negative in the stratum corneum. Compared with normal epithelia, the expression of Cx43 in dysplastic and OSCC epithelia decreased significantly. The positive rate of Cx43 in normal, mild, moderate, severe dysplasia and OSCC tissues were respectively 100.00%, 85.71%, 66.67%, 40.00%, and 33.33%. The expression of Cx43 was significantly decreased with severity increasing (P<0.05).. The expression of CK19 protein significantly increases with the development of rat tongue carcinoma, suggesting that CK19 is associated with carcinogenesis. The expression of Cx43 protein dramatically decrease with the development of rat tongue carcinoma, suggesting that the abnormal expression of Cx43 protein is associated with oral mucosa carcinoma origination. The expression of CK19 and Cx43 has negative correlation. Combined detection of CK19 and Cx43 has an important role in the early diagnosis of OSCC and can help to improve the sensitivity and specificity of the early diagnosis of OSCC.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinogenesis; Carcinoma, Squamous Cell; Connexin 43; Epithelial Cells; Epithelium; Keratin-19; Keratins; Male; Mouth Mucosa; Mouth Neoplasms; Oxides; Rats; Rats, Sprague-Dawley; Tongue; Tongue Neoplasms

Keratins play a major role in several cellular functions. Each tissue type expresses a specific set of keratins. The immense potential of keratins as diagnostic and prognostic markers for different cancers is emerging. Oral cancer is the fifteenth most common cancer worldwide. However, comprehensive information on the profile of keratins in the oral cavity is not available. Several independent reports have identified keratins using antibody based techniques which have pitfalls due to the cross reactivity of the antibodies to this set of very homologous proteins. A few recent proteomic studies have reported the identification of keratins in head and neck cancer. Majority of the studies have used tissues from the head and neck region without specifying subsites. This study reports the analysis of enriched preparations of keratins from cancer of the gingivo buccal complex (GBC) using MS, 2DE, WB, silver staining of 2DE gels and IHC. Our study reveals the absence of K4 and K13 and presence of K14, K16, and K17, in cancers of the GBC and combination of these expression patterns in the cut margins. This report also shows that K13 is glycosylated. This well characterized profile of keratins may have potential to be used in clinics.. In recent years the immense potential of keratins as diagnostic and prognostic markers for different cancers is emerging. However, comprehensive information on the profile of keratins in the oral cavity is not available. Several independent reports have identified keratins using only antibody based techniques which have pitfalls due to the cross reactivity of the antibodies to this set of very homologous proteins. This study reports the analysis of enriched preparations of keratins from a subsite of the oral cavity, the gingivo buccal complex (GBC) using mass spectrometry, 2DE, western blotting, silver staining of 2DE gels and IHC. The proteomic analysis shows the absence of K4 and K13 and presence of K14, K16, and K17 in cancers of the GBC and combination of these expression patterns in the cut margins. This well characterized profile of keratins from the gingivo buccal complex provides defined markers which may have potential to be used in the clinics.

Topics: Adult; Aged; Biomarkers; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Gingiva; Glycosylation; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Metastasis; Proteomics

Keratins are structural marker proteins with tissue specific expression; however, recent reports indicate their involvement in cancer progression. Previous study from our lab revealed deregulation of many genes related to structural molecular integrity including KRT76. Here we evaluate the role of KRT76 downregulation in oral precancer and cancer development.. We evaluated KRT76 expression by qRT-PCR in normal and tumor tissues of the oral cavity. We also analyzed K76 expression by immunohistochemistry in normal, oral precancerous lesion (OPL), oral squamous cell carcinoma (OSCC) and in hamster model of oral carcinogenesis. Further, functional implication of KRT76 loss was confirmed using KRT76-knockout (KO) mice.. We observed a strong association of reduced K76 expression with increased risk of OPL and OSCC development. The buccal epithelium of DMBA treated hamsters showed a similar trend. Oral cavity of KRT76-KO mice showed preneoplastic changes in the gingivobuccal epithelium while no pathological changes were observed in KRT76 negative tissues such as tongue.. The present study demonstrates loss of KRT76 in oral carcinogenesis. The KRT76-KO mice data underlines the potential of KRT76 being an early event although this loss is not sufficient to drive the development of oral cancers. Thus, future studies to investigate the contributing role of KRT76 in light of other tumor driving events are warranted.

Topics: Adult; Animals; Carcinogenesis; Carcinoma, Squamous Cell; Cricetinae; Disease Models, Animal; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Hyperplasia; Immunohistochemistry; Keratins; Male; Mice; Mice, Knockout; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Grading; Neoplasm Staging; Prognosis; Reproducibility of Results

The study was aimed at analysing and identifying the proteins that are differentially expressed in oral squamous cell carcinoma (OSCC) compared to adjacent non-tumour tissue.. Two-dimensional (2D) sodium dodecyl sulphate-polyacrylamide gel electrophoresis accompanied by mass spectrometry (matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry and liquid chromatography-tandem mass spectrometry) was used to analyse and identify the differentially expressed proteins in 10 pairs of tumours and adjacent non-tumour tissues from five cases of early-stage and five cases of late-stage OSCC. The statistical differences of the protein spots were analysed by the Wilcoxon signed-rank test. A validation study using immunohistochemistry and quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was performed.. A total of 68 proteins (63 up-regulated, five down-regulated) were differentially expressed in early-stage disease, and 39 proteins (37 up-regulated, two down-regulated) were significantly altered in late-stage disease. Among these, 14 proteins were altered in both groups. A total of 44 proteins were identified, including heat shock proteins (HSPs: Hsp90, HSPA5 and HSPA8), keratins (K1, K6A and K17), tubulin, cofilin 1, 14-3-3σ and metabolic enzymes. These proteins are involved in various cellular processes essential for cell growth, survival and cell migration. The validation study on α-tubulin and 14-3-3σ using immunohistochemistry and KIAA1199 expression using real-time RT-PCR confirmed the results in proteomics analysis.. The study identified many proteins, both known and unknown, for cancer cell processes. At least two proteins, KIAA1199 and Horf6, are novel for oral cancer.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Down-Regulation; Electrophoresis, Gel, Two-Dimensional; Endoplasmic Reticulum Chaperone BiP; Gene Expression Regulation, Neoplastic; Heat-Shock Proteins; Humans; Hyaluronoglucosaminidase; Immunohistochemistry; Keratins; Mouth Neoplasms; Peroxiredoxin VI; Proteins; Proteomics; Real-Time Polymerase Chain Reaction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thailand; Tubulin; Up-Regulation

Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine with pro-inflammatory functions and involved in tumorigenesis. The aim of this study was to evaluate the expression and localization of the macrophage MIF in oral squamous carcinoma (OSC). In addition, the relationship between MIF expression and clinicopathological parameters such as survival data, tobacco use, alcohol habits, TNM stage, tumor graduation, and peritumoral inflammatory infiltrate were evaluated.. Using immunohistochemistry, expression and localization of MIF was detected in 44 specimens of OSC. The absolute number and relative proportions of MIF-positive cells detected were also determined separately for tumor parenchyma vs. stroma. All counts were determined from 10 consecutive high-power fields using an integration graticule. Moreover, some parameters were analyzed separately for lip and intra-oral cancers.. Migration inhibitory factor-positive cells were observed in both the tumor parenchyma and in inflammatory cells of all specimens. In contrast, MIF expression was not detected in tumoral nests associated with poorly differentiated tumors. In specimens of lip cancer, a greater number of MIF-positive stromal immune cells were detected than in intra-oral cancer specimens (Mann-Whitney test, P = 0.049).. Oral squamous carcinoma cells consistently express MIF independent of their location. Lip tumors presented more MIF-positive peritumoral inflammatory cells, similar to control, suggesting that immunological differences in leukocyte activation exist between in lip and intra-oral cancers.

Topics: Alcohol Drinking; Carcinoma, Squamous Cell; Cell Count; Cohort Studies; Epithelium; Female; Humans; Inflammation; Keratins; Leukocytes; Leukoplakia, Oral; Lip Neoplasms; Macrophage Migration-Inhibitory Factors; Male; Middle Aged; Mouth Neoplasms; Neoplasm Grading; Neoplasm Staging; Smoking; Stromal Cells; Survival Rate

To determine the feasibility of viable storage of head and neck squamous cell carcinoma (HNSCC) for regrowth of cells in culture.. Laboratory-based translational study.. Methods for intermediate-term frozen storage of viable HNSCC were explored using small pieces of primary tumor and dissociated HNSCC cells after short-term culture. Viable cells after freezing were confirmed by adherence to tissue culture plates, cell morphology, and increased cell or colony density. Two cultures were immunostained for cytokeratin to confirm epithelial origin of viable cultured cells after freezing.. Six primary HNSCCs (two oral cavity, three larynx, one oropharynx) and two HNSCCs that had been passaged through a xenograft (two oral cavity) were dissociated to single cells and grown in short-term cell culture for 0 to 12 passages. After short-term culture, cells were frozen for up to 8 months, thawed, and replated. Frozen cells derived from all tumors (six primary and two xenografts) were successfully replated with cultures lasting >7 days with seven of eight tumors presenting increased colony or cell density over 1 week of growth after freezing. In total, 15 of 15 tested samples derived from six primary and two xenografted HNSCCs were viable after freezing.. In the current study, we show that biopreservation of primary or xenografted HNSCC using short-term cell culture is feasible. Initial short-term cell culture was required for successful storage and viability of frozen cells. These proof-of-principle studies, if more widely implemented, could improve preclinical testing of new therapies for HNSCC.

Topics: Carcinoma, Squamous Cell; Cryopreservation; Feasibility Studies; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Mouth Neoplasms; Oropharyngeal Neoplasms; Squamous Cell Carcinoma of Head and Neck; Tissue Banks; Transplantation, Heterologous; Tumor Cells, Cultured

Basaloid squamous cell carcinoma (BSCC) is rare variant of oral squamous cell carcinoma (OSCC) with predilection for upper aerodigestive tract. Although it is characterized by distinct histologic features it is often confused with conventional OSCC and other basaloid tumors. The study aims to establish differentiating features of BSCC with oral basaloid tumors using immunohistochemical (IHC) markers. This retrospective study included 34 cases, including BSCC, OSCC, and basaloid tumors. IHC staining was performed with primary antibodies against cytokeratin (CK) 19, 14, 8/18, α-smooth muscle actin (αSMA), p53, and MMP-9. A prominent CK 19, 14, and 8/18 expression was observed in BSCC as compared with basaloid tumors suggesting of basal cell origin with undifferentiated type of tumor cells. Expression of αSMA was intense in tumor cells of myoepithelial differentiation but lacked in BSCC. The intense expression of p53 and MMP-9 was noted in all basaloid malignancies. Considering standard histologic criteria in diagnosing BSCC, when in confusion with other basaloid tumors, IHC markers gain importance. Hence, enhanced expression of CK 19, 8/18, and 14 and coexistence of p53 and MMP-9 expression and negativity for αSMA suggest an accurate diagnosis of BSCC.

Topics: Actins; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Male; Matrix Metalloproteinase 9; Middle Aged; Mouth Neoplasms; Neoplasms, Basal Cell; Tumor Suppressor Protein p53

Topics: Adult; Aged; Carcinoma, Squamous Cell; Epithelial Cells; Female; Follow-Up Studies; gp100 Melanoma Antigen; Humans; Keratins; Lymphatic Metastasis; Male; MART-1 Antigen; Melanins; Melanocytes; Melanoma-Specific Antigens; Middle Aged; Mouth Floor; Mouth Mucosa; Mouth Neoplasms; S100 Proteins; Tongue Neoplasms

Keratins are intermediate filament family proteins which are predominantly expressed in the epithelial cells. Most of the studies which evaluate the status of keratins in clinical samples of the oral cavity are based on the identification of their presence and localization by immunohistochemistry using monoclonal antibodies. It is very well known that many monoclonal/polyclonal antibodies show cross-reactivity with the other closely related or non-related proteins. This cross-reactivity might be the result of epitope similarity, but it is not always necessary. Therefore studies done with only antibody based techniques can mislead interpretation unless they are validated with additional techniques like mass-spectrometry. In this investigation we have evaluated the status of keratin 18 in cancer of buccal mucosa using 1DE, 2DE and western blotting with monoclonal antibody to keratin 18. The patterns emerging showed aberrant as well as differential expression of K18 in adjacent normal versus tumor tissue samples of buccal mucosa. Mass spectrometry analysis of the immunodetected spots however revealed that it is keratin 13. Thus this study emphasizes the necessity of validation of antibody based findings when dealing with proteins of a large family having similarity/homology in amino acid sequence.

Topics: Amino Acid Sequence; Antibodies; Antibody Specificity; Carcinoma; Cross Reactions; False Positive Reactions; Humans; Immunohistochemistry; Keratins; Mass Spectrometry; Microdissection; Mouth Neoplasms; Sensitivity and Specificity; Tissue Extracts

Topics: Carcinoma, Adenosquamous; Epithelium; Female; Humans; Immunohistochemistry; Keratins; Middle Aged; Mouth Mucosa; Mouth Neoplasms

P-cadherin belongs to the family of Ca(2+)-dependent homophilic glycosylated cell adhesion molecules. In the normal oral epithelium it shows a strong expression in the basal cell layer which gradually decreases in the suprabasal cell layers. The exact role of P-cadherin during the development and homeostasis of the oral epithelium has not been elucidated, yet. Here, we show for the first time that P-cadherin controls differentiation by regulating cytokeratin (CK) 1/10 expression in primary oral keratinocytes (POK) from normal, but interestingly not in POKs from oral squamous cell carcinoma (OSCC) tissue. SiRNA knockdown of P-cadherin in normal POKs revealed a strong upregulation of CK1/10 expression on mRNA and protein level. In contrast, E-cadherin knockdown in normal oral keratinocytes did not show any influence on CK1/10 expression. Moreover, in comparison with normal control keratinocytes normal oral keratinocytes with reduced P-cadherin expression displayed an enhanced expression and a stronger nuclear staining of C/EBP-beta, a well-known regulator of CK1/10 expression in keratinocytes. Furthermore, after P-cadherin knockdown in normal POKs the promoter activity of a C/EBP-responsive luciferase construct was significantly higher than in normal POKs with regular P-cadherin expression. Additionally, we noticed a proliferation advantage in normal oral keratinocytes in contrast to keratinocytes with diminished P-cadherin expression. However, the inverted effect was seen in tumor derived primary oral keratinocytes. In summary, we show that P-cadherin contributes to the keratinocyte differentiation in the oral epithelium by influencing the CK1 and CK10 expression via C/EBP-beta-mediated signaling in normal but not in tumor derived oral keratinocytes from OSCC patients.

Topics: Cadherins; Carcinoma, Squamous Cell; CCAAT-Enhancer-Binding Protein-beta; Cell Differentiation; Cell Line; Cell Line, Tumor; Cell Proliferation; Humans; Keratinocytes; Keratins; Mouth Mucosa; Mouth Neoplasms; Promoter Regions, Genetic; RNA, Messenger; RNA, Small Interfering; Signal Transduction; Up-Regulation

Topics: Airway Obstruction; Dermoid Cyst; Diagnosis, Differential; Epithelium; Humans; Infant; Keratins; Magnetic Resonance Imaging; Male; Mouth Floor; Mouth Neoplasms; Sebaceous Glands

Darier's disease (DD) is an inherited autosomal-dominant skin disorder characterized histologically by loss of adhesion between keratinocytes. DD is typically caused by mutations in sarcoendoplasmic reticulum Ca(2+)-ATPase isoform 2 (SERCA2), a major regulator of intracellular Ca(2+) homeostasis in the skin. However, a defined role for SERCA2 in regulating intercellular adhesion remains poorly understood. We found that diminution of SERCA2 function by pharmacological inhibition or siRNA silencing in multiple human epidermal-derived cell lines was sufficient to disrupt desmosome assembly and weaken intercellular adhesive strength. Specifically, SERCA2-deficient cells exhibited up to a 60% reduction in border translocation of desmoplakin (DP), the desmosomal cytolinker protein necessary for intermediate filament (IF) anchorage to sites of robust cell-cell adhesion. In addition, loss of SERCA2 impaired the membrane translocation of protein kinase C α (PKCα), a known regulator of DP-IF association and desmosome assembly, to the plasma membrane by up to 70%. Exogenous activation of PKCα in SERCA2-deficient cells was sufficient to rescue the defective DP localization, desmosome assembly, and intercellular adhesive strength to levels comparable to controls. Our findings indicate that SERCA2-deficiency is sufficient to impede desmosome assembly and weaken intercellular adhesive strength via a PKCα-dependent mechanism, implicating SERCA2 as a novel regulator of PKCα signaling.

Topics: Calcium; Carcinoma, Squamous Cell; Cell Adhesion; Cell Communication; Cell Line, Tumor; Darier Disease; Desmoplakins; Desmosomes; Humans; Intermediate Filaments; Keratins; Mouth Neoplasms; Protein Kinase C-alpha; RNA, Small Interfering; Sarcoplasmic Reticulum Calcium-Transporting ATPases; Signal Transduction

Assessing epithelial dysplasia to predict malignant transformation remains problematic in many tissues because grading systems are poorly structured and individual features poorly defined. Dysplasia grading is criticised for lack of reproducibility and poor predictive value. Grading systems for upper aerodigestive tract dysplasia have evolved over several decades and are not supported by good outcome experimental data..  This study analysed the individual features of dysplasia in 86 oral dysplastic lesions and determined the reproducibility of scoring for each, and correlated them with other features and clinical factors using complex clustering analyses.. A uniform pattern of dysplasia was found in 37 lesions, focal dysplasia in 36 and in 13 lesions dysplasia formed complex discontinuous patterns. There was wide variation in reproducibility of scoring of individual features and many, including thickness, some types of rete morphology, basaloid cell anisonucleosis, basal dyscohesion, and dyskeratosis as deep single cells correlated with sub-sites. Rete morphology, type of keratinisation, hyperchromatism of the basaloid compartment, prickle cell anisonucleosis and extension down salivary ducts correlated with smoking. Conventional grading and oral intraepithelial neoplasia (OIN) grading by 'thirds affected' showed strong correlation overall but scores obtained with the OIN system tended to a higher grade at all sites except soft palate/fauces. There was poor correlation between the systems for moderate dysplasia and also severe dysplasia at some sites. Individual features could not be shown to cluster to form distinct patterns of dysplasia.. These variations may account in part for the lack of reproducibility and poor predictive value of the grading systems in current use and could inform the design of future grading systems.

Topics: Carcinoma in Situ; Cell Adhesion; Cell Nucleus; Cell Transformation, Neoplastic; Chromatin; Epithelial Cells; Epithelium; Female; Humans; Keratins; Leukoplakia, Oral; Lip Neoplasms; Male; Mitosis; Mouth Floor; Mouth Mucosa; Mouth Neoplasms; Neoplasm Grading; Palatal Neoplasms; Palate, Soft; Precancerous Conditions; Reproducibility of Results; Salivary Ducts; Tongue Neoplasms

Local failure occurs in 13.9-62.6% and it is a well known indicator of poor prognosis in patients with oral squamous cell carcinoma (OSCC), despite aggressive treatments. The purpose of this study was to investigate the value of histopathology and molecular biomarkers in predicting the development of early local recurrence.. This study included a total of 69 patients. There were 23 patients with early recurrent OSCC and 46 patients without local recurrence with the same clinical stage and tumor site, in a pair-matched study design. Their charts were retrospectively analyzed. All surgical specimens of the primary tumors were evaluated according to the system proposed by Anneroth et al. and immunohistochemical for ErbB2 and FAS were performed.. A significant correlation of early local recurrence with grade of histological malignancy (more than 15 points) was observed (Fisher's exact test, P = 0.03). Early local recurrence was also significantly associated with weak FAS expression and strong intracytoplasmic ErbB2 staining (Mantel-Haenszal chi-square, P = 0.0038 and P = 0.0068, respectively). Histological grade of malignancy (more than 15 points) was also correlated with reduced survival (log-rank, P = 0.06). Among the histopathological parameters, keratinization, pattern of invasion and inflammation were important for overall survival (log-rank, P < 0.0001). Regarding the biomarkers, only FAS was significantly associated with overall survival (log-rank, P = 0.0002). Moreover, a positive correlation of FAS and membrane ErbB2 expression with keratinization was noticed.. Histopathological characteristics and the expression of FAS and ErbB2 carry prognosis importance in local recurrence and overall survival in OSCC.

Topics: Biomarkers, Tumor; Case-Control Studies; Cell Membrane; Cytoplasm; fas Receptor; Female; Follow-Up Studies; Forecasting; Gene Expression Regulation, Neoplastic; Genes, erbB-2; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasm Staging; Receptor, ErbB-2; Retrospective Studies; Survival Rate

We compared findings of optical coherence tomography (OCT) with histopathological results of suspicious oral lesions to assess the feasibility of using OCT to identify malignant tissue. Thirty-four oral lesions from 27 patients had swept-source frequency-domain OCT. Four variables were assessed (changes in keratin, epithelial, and sub-epithelial layers, and identification of the basement membrane) and from this we calculated whether or not there were architectural changes. These data were then compared with histopathological results. Two clinicians, who were unaware of the clinical and histopathological diagnoses, decided whether biopsy was necessary. The basement membrane was recognised in only 15 oral lesions. OCT could identify diseased areas but could not provide a diagnosis or differentiate between lesions. The two clinicians, who recommended biopsy agreed in all cases. This pilot study confirms the feasibility of using OCT to identify architectural changes in malignant tissues.

Topics: Basement Membrane; Biopsy; Carcinoma, Squamous Cell; Diagnosis, Differential; Epithelium; Erythroplasia; Feasibility Studies; Humans; Image Processing, Computer-Assisted; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Oral Ulcer; Pilot Projects; Precancerous Conditions; Tomography, Optical Coherence; Tongue Neoplasms

Oral spindle cell carcinoma (SpCC) is a rare variant of oral squamous cell carcinoma (SCC). The aims of this study were to compare the clinicopathologic and immunohistochemical features of oral SpCC with conventional oral SCC.. Five cases of oral SpCC and 10 cases of oral SCC (five well-differentiated and five poorly differentiated) were evaluated through conventional hematoxylin and eosin staining and immunohistochemical reactions to cytokeratins (CK), vimentin, desmin, smooth muscle actin, muscle-specific actin, S-100 protein, epithelial membrane antigen (EMA), p53, and ki-67.. Oral SpCC showed predilection for males on their sixth decade of life, presenting clinically as painful infiltrative ulcers or ulcerated exophytic polypoid masses, preferably located on the alveolar mucosa. Mesenchymal markers were expressed in the spindle cell but not in the carcinomatous component of SpCC, and it was negative in all SCC. CKs AE1/AE3, 6, 14, and EMA were positive on both carcinomatous and spindle cell components of most SpCCs. These tumors also presented higher p53 and ki-67 expression and no CK 1 expression in contrast to well-differentiated SCC.. Oral SpCC presented a different clinical profile than conventional SCC and histopathologic features and p53 and ki-67 expression closer to poorly differentiated SCC. Besides mesenchymal markers, CK AE1/AE3, 6, 14, and EMA expression on spindle cells may be useful as an adjunct on microscopical differential diagnosis of SpCC.

Topics: Actins; Adult; Age Factors; Aged; Carcinoma; Carcinoma, Squamous Cell; Desmin; Female; Humans; Immunohistochemistry; Keratin-13; Keratin-14; Keratin-6; Keratin-8; Keratins; Ki-67 Antigen; Male; Middle Aged; Mouth Neoplasms; Mucin-1; S100 Proteins; Sex Factors; Tumor Suppressor Protein p53; Vimentin

Metastatic clear cell renal cell carcinoma (CCRCC) should be considered in differential diagnosis of intraoral clear cell tumors, including mucoepidermoid carcinoma (MEC).. We compared the clinical, histologic, histochemical, and immunohistochemical characteristics of 9 oral metastatic CCRCCs and 8 intraoral clear cell MECs.. Oral metastatic CCRCC affected salivary-gland containing tissues in 7 cases (78%). Microscopically, oral metastasis revealed a proliferation of neoplastic clear cells arranged in an alveolar pattern with central blood vessels, features that were not seen in any intraoral clear cell MEC. Mucicarmine staining was positive only in clear cell MEC. Immunohistochemistry showed similarities in cytokeratin expression; vimentin and CD10 were expressed in all oral metastatic CCRCCs but in only 1 clear cell MEC each.. Besides clinical history, the alveolar pattern, vessel distribution, absence of mucicarmine staining, and vimentin and CD10 immunoexpression are useful in histologic differential diagnosis of CCRCC and clear cell MEC.

Topics: Adenocarcinoma, Clear Cell; Adult; Aged; Aged, 80 and over; Carcinoma, Mucoepidermoid; Carcinoma, Renal Cell; Carmine; Cell Nucleus; Coloring Agents; Cytoplasm; Diagnosis, Differential; Female; Hemorrhage; Histocytochemistry; Humans; Immunohistochemistry; Keratins; Male; Microvessels; Middle Aged; Mouth Neoplasms; Neprilysin; Salivary Gland Neoplasms; Vimentin

This study compares synchronous oral squamous cell carcinomas (OSCCs) with single primary OSCCs to assess the histopathologic parameters with a known prognostic significance.. Twenty-eight cases of synchronous OSCCs and a control group of single primary OSCCs were compared for 15 histologic prognostic variables.. Results showed significantly less amount of abnormal mitoses (synchronous-1: P = .002; synchronous-2: P = .006) and tumor-induced stroma (synchronous-1: P = .011; synchronous-2: P = .001) in synchronous OSCCs than in single primary OSCCs. Depth of invasion was considerably lower in synchronous OSCCs than in single primary OSCCs (synchronous-1: P = .007; synchronous-2: P = .002). Lymph node metastasis (synchronous-1: P = .051; synchronous-2: P = .051) was found to be rare in synchronous OSCCs compared with single primary OSCCs.. Synchronous OSCCs show less aggressive histopathologic features than single primary OSCCs.

Topics: Aged; Carcinoma, Squamous Cell; Case-Control Studies; Cell Differentiation; Female; Gingival Neoplasms; Humans; Keratins; Lip Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Mitosis; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasms, Multiple Primary; Palatal Neoplasms; Tongue Neoplasms

The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression.. The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells.. We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (ARID4A, CALR, GNB2L1, RNF10, SQSTM1, USP9X) were validated by real time PCR.. A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.

Topics: Annexin A5; Apoptosis; Cell Proliferation; Down-Regulation; Electrophoresis, Gel, Two-Dimensional; Fibroblasts; Gene Expression Regulation, Neoplastic; Genomics; Hep G2 Cells; Humans; Keratins; Mouth Neoplasms; Nucleic Acid Hybridization; Proteome; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stromal Cells; Vimentin

Met, the hepatocyte growth factor receptor, is important in transducing signals for tumour growth and metastasis. The aim of this study was to examine the pattern of Met expression and its value as a prognostic factor in oral squamous cell carcinomas (OSCCs). The material consisted of 53 OSCCs and five healthy controls from normal oral mucosa supplied with cell lines, 10 organotypic models supplied with oral cancer cells, and three organotypic models supplied with normal keratinocytes. Met protein expression was assessed by immunohistochemistry and western blotting. Met expression was scarce and limited to the basal layer in normal oral mucosa, but was more extensive in the tumours. Cytoplasmic expression of Met was found in the majority of the tumours, and nuclear expression was found in 72%, including a high fraction of the cells located at the invasive front. Organotypic models with normal or malignant oral cells yielded principally similar results as in the mucosa and the cancers, respectively. A smaller amount of Met immunoreactivity was detected, by western blotting, in the nuclear fraction of cultured oral cancer cells. In conclusion, Met was upregulated in OSCCs and was also found in the nucleus. However, Met was not a marker for prognosis in this study.

Topics: Adenocarcinoma; Biomarkers, Tumor; Breast Neoplasms; Carcinoma; Carcinoma, Squamous Cell; Cell Culture Techniques; Cell Line; Cell Nucleus; Cells, Cultured; Coculture Techniques; Cytoplasm; Female; Fibroblasts; Gingiva; Humans; Keratinocytes; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness; Prognosis; Proto-Oncogene Proteins c-met; Skin Neoplasms; Tissue Scaffolds; Tongue Neoplasms; Up-Regulation

Oral squamous cell carcinoma (SCC) is an aggressive tumor with a poor 5-year survival rate. Oral SCC can undergo epithelial to mesenchymal transition (EMT). We previously showed that the epithelial integrin alphavbeta6 complexes with Fyn kinase in oral SCC to promote EMT. Using immunofluorescence microscopy and Western blotting, we evaluated whether the expression of specific markers of EMT were influenced by modulating serum concentration (ie. growth factors). The SCC cultures were grown under contrasting levels of serum. In low serum (1%), Fyn promoted EMT; whereas suppression of Fyn kinase promoted the epithelial phenotype. However, when the SCC cells were grown in 10% serum, activation of Fyn had the reverse effect. Lastly, cell migration was evaluated under low serum conditions (1% FBS). Activation of Fyn promoted SCC cell migration and its suppression thwarted SCC migration toward FN. These results indicate that the activation of Fyn kinase as well as local growth factor concentration modulate EMT in oral SCC.

Topics: Blotting, Western; Cadherins; Carcinoma, Squamous Cell; Cell Communication; Cell Movement; Culture Media; Enzyme Activation; Epithelial Cells; Humans; Keratins; Mesoderm; Microscopy, Fluorescence; Mouth Neoplasms; Proto-Oncogene Proteins c-fyn; Serum; Signal Transduction

To investigate the role of the Chinese herbal medicine Xianhuayin on the reversal of 7,12-dimethylbenz[a]anthracene (DMBA)-induced premalignant mucosal lesions in the oral buccal pouch of golden hamsters.. The animals were randomly divided into a non-diseased control group (n=5) and an experimental group including 50 animals in which the buccal mucosa had been painted with DMBA (0.5% in acetone) to generate an oral mucosa premalignant lesion. Animals in the experimental group were further divided into Xianhuayin-treated group (n=30), untreated premalignant lesion group (n=10) and normal saline (NS)-treated group (n=10). The cheek (buccal) pouch mucosa of the golden hamsters in each group was observed with light and electron microscopy eight weeks after intragastric administration with NS or Xianhuayin.. In the non-diseased control group, the buccal mucosa was keratinized and stratified squamous epithelium under a light microscope. In the untreated premalignant lesion group, variable degrees of epithelial dysplasia was observed. The irregular epithelial mucosa gradually became distinct in the Xianhuayin-treated group. Scanning electronic microscopic (SEM) analysis showed that surface of the cells exhibited honeycomb structures in the hamster of untreated-group. The cells were morphologically irregular, overlapped and loosened in the untreated premalignant lesion group. Most of the cell surface exhibited honeycomb structure in the Xianhuayin-treated group. Transmission electronic microscopic (TEM) analysis showed that buccal mucosal epithelial cells were morphologically regular in the non-diseased control group. Desmosomes and tonofibrils were reduced and the nucleus was morphologically irregular in the untreated premalignant lesion group. In the Xianhuayin-treated group, the widening intercellular gap was gradually reduced, desmosomes and the cells becoming morphologically regular. No significant difference was observed between the hamsters in NS-treated group and those in the untreated premalignant lesion group. Significant therapeutic efficacy was observed in the group receiving Xianhuayin.. Xianhuayin is effective in the reversal of DMBA-induced premalignant lesions in the buccal pouch of golden hamsters.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Amomum; Animals; Anticarcinogenic Agents; Carcinogens; Carthamus tinctorius; Cell Nucleus; Cricetinae; Desmosomes; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial Cells; Epithelium; Glycyrrhiza; Hyperplasia; Intercellular Junctions; Intermediate Filaments; Keratins; Mesocricetus; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Mouth Mucosa; Mouth Neoplasms; Philodendron; Poria; Precancerous Conditions; Random Allocation; Sodium Chloride

The present study aimed to investigate the membrane stabilizing effect of Thymoquinone (TQ) on cell surface glycoconjugates and cytokeratin expression against DMBA induced hamster buccal pouch carcinogenesis. 0.5% DMBA painting (three times per week) in hamster buccal pouches for 14 weeks resulted in the formation of well developed oral squamous cell carcinoma. We observed 100% tumor formation with marked abnormalities of glycoconjugates status in tumor bearing hamsters as compared to control animals. Oral administration of TQ at a dose of 30 mg/kg body weight, to DMBA painted hamsters on alternate days for 14 weeks, reduced the tumor formation as well as protected the levels of cell surface glycoconjugates in DMBA painted hamsters. The present study thus suggests that TQ has potent chemopreventive efficacy as well as protected the abnormalities on cell surface glycoconjugates during DMBA induced hamster buccal pouch carcinogenesis.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Antineoplastic Agents; Benzoquinones; Carcinogens; Carcinoma, Squamous Cell; Chemoprevention; Cricetinae; Drug Evaluation, Preclinical; Glycoconjugates; Keratins; Male; Membrane Glycoproteins; Mesocricetus; Mouth Neoplasms; Neoplasms, Experimental

Oral verrucous hyperplasia (OVH) and oral erythroleukoplakia (OEL) are two oral precancerous lesions with relatively high malignant transformation potential. One of the best cancer prevention strategies is to use a conservative and effective treatment modality to eliminate oral precancers to stop their further malignant transformation. Our previous studies have shown that the topical 5-aminolevulinic acid-mediated photodynamic therapy (topical ALA-PDT) using the 635-nm light-emitting diode (LED) light is very effective for OVH and OEL lesions.. Because the laser machine is a more-popular light source than the LED device in PDT clinics, in this study 40 OVH and 40 OEL lesions were treated once a week with the same PDT protocol but using the 635-nm laser light to evaluate whether this laser light-mediated topical ALA-PDT was also effective for OVH and OEL lesions.. We found that all the 40 OVH lesions exhibited complete response (CR) after an average of 3.6 PDT treatments. Of the 40 OEL lesions, 38 showed CR after an average of 3.4 PDT treatments and two showed partial response (PR). Better PDT outcomes were significantly associated with OVH and OEL lesions with the smaller size, pink to red color, epithelial dysplasia, or thinner surface keratin layer.. This study indicates that the laser light-mediated topical ALA-PDT is also very effective for OVH and OEL lesions. Therefore, we suggest that topical ALA-PDT using either the LED or laser light may serve as the first-line treatment of choice for OVH and OEL lesions.

Topics: Adult; Aged; Aged, 80 and over; Aminolevulinic Acid; Biopsy; Carcinoma in Situ; Erythroplasia; Female; Humans; Hyperplasia; Keratins; Leukoplakia, Oral; Low-Level Light Therapy; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Photochemotherapy; Photosensitizing Agents; Precancerous Conditions; Remission Induction; Treatment Outcome

Snail is a regulator of epithelial-mesenchymal transition (EMT) and considered crucial to carcinoma metastasis, myofibroblast transdifferentiation, and fibroblast activation. To investigate the role of Snail in oral squamous cell carcinoma (OSCC), its immunohistochemical expression was analysed in 129 OSCC samples and correlated to nodal metastasis, histological grade, E-cadherin, and alpha smooth-muscle-actin (alpha SMA). The results were compared to findings in 23 basal cell carcinomas (BCC). Additionally, the influence of TGF beta 1 and EGF on Snail, E-cadherin, vimentin, and alpha SMA expression was analysed in two OSCC cell lines. As a result, Snail-positive cells were mainly found in the stroma of the OSCC invasive front without statistically significant correlation to histological grade or nodal metastasis. Snail was co-localised to alpha SMA but not to E-cadherin or cytokeratin and showed a significant correlation to the loss of membranous E-cadherin. All BCCs were Snail negative. In OSCC culture, the growth-factor-mediated EMT-like phenomenon was accompanied by alpha SMA down-regulation. In summary, Snail expression in OSCC is a stromal phenomenon associated with the myofibroblast phenotype and not related to growth-factor-mediated transdifferentiation of the carcinoma cells themselves. Consequently, Snail immunohistochemistry cannot contribute to the prediction of the metastatic potential. Furthermore, stromal Snail expression is suggested to be the result of mutual paracrine interaction of fibro-/myofibroblasts and dedifferentiated carcinoma cells leading to the generation of a special type of carcinoma-associated fibroblasts.

Topics: Actins; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cadherins; Carcinoma, Squamous Cell; Epidermal Growth Factor; Fibroblasts; Humans; Keratins; Middle Aged; Mouth Neoplasms; Myoblasts; Snail Family Transcription Factors; Transcription Factors; Transforming Growth Factor beta1; Vimentin

The purpose of this report is to present the clinical and histological features of a basaloid squamous cell carcinoma (BSCC) occurring in the retromolar trigone of a 59-year-old man and to relate its immunohistochemical characteristics.. BSCC is an aggressive distinct variant of squamous cell carcinoma (SCC) requiring recognition as a separate entity from SCC due to its peculiar behavior.. A clinical examination revealed a 12x07x07 mm nodular mass with a rubbery consistency, defined borders, covered by reddish mucosa and an absence of bleeding upon palpation. Histologically, nests and cords of closely packed, moderately pleomorphic basaloid cells with nuclear palisading along the periphery of the neoplastic nests surrounded by a fibrous stroma were found.. Since this tumor can mimic other neoplasms such as adenoid cystic carcinoma, neuroendocrine carcinoma, and basal cell adenocarcinoma, histological features are essential to differentiate between them. Furthermore, immunohistochemical testing can provide valuable diagnostic information that can have a profound impact on treatment options and the prognosis.. BSCC needs to be differentiated from other neoplasms as early as possible because of its adverse prognosis. Clinicians are advised to conduct a mucosal evaluation during oral examinations and take a thorough medical history which could ultimately save the life of a patient.

Topics: Adenocarcinoma; Carcinoma, Adenoid Cystic; Carcinoma, Neuroendocrine; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cell Nucleus; Diagnosis, Differential; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Laminin; Male; Middle Aged; Mitosis; Mouth Neoplasms; Necrosis; Tumor Suppressor Protein p53

Pathologists having drawn attention to the morphological and functional characteristics of invasive tumor front (ITF), we attempted to investigate the difference of E-cadherin expression between the ITF and center/superficial part as well as the prognostic value of E-cadherin expression at the ITF.. We performed a retrospective analysis of the correlation between E-cadherin protein expression at the ITF and center/superficial part and prognostic factors. We also detected the difference of E-cadherin expression at the ITF and center/superficial part using immunohistochemical method, which was further supported in 20 fresh OSCC samples by confocal laser microscopy and RT-PCR methods.. The expression of E-cadherin in the same tumor was heterogeneous. Both protein (t test, P

Topics: Adult; Aged; Aged, 80 and over; Cadherins; Carcinoma, Squamous Cell; Cell Nucleus; Female; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Keratins; Male; Microscopy, Confocal; Middle Aged; Mouth Neoplasms; Neoplasm Invasiveness; Prognosis; Proportional Hazards Models; Retrospective Studies; Reverse Transcriptase Polymerase Chain Reaction; Young Adult

In this study, we evaluated whether the forced expression of beta6 integrin would modulate the epithelial to mesenchymal transition (EMT). When the full length beta6 integrin was expressed in poorly invasive squamous cell carcinoma SCC9 cells, the resulting SCC9/6 cells acquired a fibroblast-like morphology, increased expression of the mesenchymal marker vimentin and reduced expression of the epithelial markers keratin and E-cadherin. SCC9beta6D1 cells, which express a truncated form of beta6 subunit lacking the C-terminal 11 amino acids (AA), retained their epithelial morphology and did not alter vimentin or E-cadherin expression. This suggests that the full-length beta6 subunit can induce EMT in oral SCC cells. We previously showed that expression of beta6 increases both MMP-3 activation and tenascin-C expression and we now show that both molecules are MEK dependent. These results also demonstrate that the terminal 11 AA of beta6 contain information important for establishing an epithelial to mesenchymal transition.

Topics: Antigens, Neoplasm; Cadherins; Carcinoma, Squamous Cell; Cell Line, Tumor; Enzyme Activation; Epithelial Cells; Humans; Integrins; Keratins; MAP Kinase Kinase Kinases; Matrix Metalloproteinase 3; Matrix Metalloproteinase Inhibitors; Mesoderm; Mouth Neoplasms; Tenascin; Vimentin

The chemopreventive efficacy of turmeric has been established in experimental systems. However, its mechanism(s) of action are not fully elucidated in vivo. The present study investigates the mechanism of turmeric-mediated chemoprevention in 7,12-dimethylbenz(a)anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis at 2, 4, 6, 10 and 12 weeks. Dietary turmeric (1%) led to decrease in DMBA-induced tumor burden and multiplicity, and enhanced the latency period in parallel, to its modulatory effects on oncogene products and various cellular responses during HBP tumorigenesis. DMBA-induced expression of ras oncogene product, p21 and downstream target, the mitogen-activated protein kinases were significantly decreased by turmeric during HBP carcinogenesis. Turmeric also diminished the DMBA-induced mRNA expression of proto-oncogenes (c-jun, c-fos) and NF-kappaB, leading to decreased protein levels and in further attenuation of DMBA-induced AP-1/NF-kappaB DNA-binding in the buccal pouch nuclear extracts. Besides, buccal pouch of hamsters receiving turmeric diet showed significant alterations in DMBA-induced effects: (a) decrease in cell proliferation (diminished PCNA and Bcl2 expression), (b) enhanced apoptosis (increased expression of Bax, caspase-3 and apoptotic index), (c) decrease in inflammation (levels of Cox-2, the downstream target of AP-1/NF-kappaB, and PGE2) and (d) aberrant expression of differentiation markers, the cytokeratins (1, 5, 8, and 18). Together, the protective effects of dietary turmeric converge on augmenting apoptosis of the initiated cells and decreasing cell proliferation in DMBA-treated animals, which in turn, is reflected in decreased tumor burden, multiplicity and enhanced latency period. Some of these biomarkers are likely to be helpful in monitoring clinical trials and evaluating drug effect measurements.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Apoptosis; Cell Proliferation; Cheek; Cricetinae; Curcuma; Diet; Keratins; Male; MAP Kinase Signaling System; Mesocricetus; Mouth Neoplasms; NF-kappa B; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins p21(ras); Transcription Factor AP-1

To investigate the clinical application value of serum tumor markers detection combined with support vector machine (SVM) model in the diagnosis of oral squamous cell carcinoma.. Serum levels of neuron-specific enolase (NSE), cancer antigen 242 (CA242), cancer antigen 19-9 (CA199), carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA), cancer antigen 72-4 (CA724), cancer antigen 21-1 (CA211) and alpha fetoprotein (AFP) were detected with enzyme-linked immunosorbent assay (ELISA) and time-resolved fluoroimmunoassay (TRFIA) in 163 oral squamous cell carcinoma patients and 160 healthy persons. All the data was analyzed with SVM; the SVM models for diagnosis of oral squamous cell carcinoma were created, trained and validated by cross validation.. Among the 163 oral squamous cell carcinoma patients, there were 128 males and 35 females with the male-to-female ratio of 3.66:1; the age ranged from 30 to 85 years old with a mean age of 59.3 years old; according to the primary site of tumor, 72 cases in tongue, 34 in gingiva, 22 in buccal mucosa, 15 in palatal mucosa, 13 in floor of mouth, 4 in lip and 3 in retromolar region; according to the TNM-UICC classification, there were 33 patients at stage T1, 72 at T2, 44 at T3, 14 at T4, 119 at N0, 42 at N1, 2 at N2, 159 at M0, 4 at M1, 27 at clinical stage I, 51 at stage II, 52 at III, and 33 at IV; according to the pathological differentiation grade, 109 tumors were well differentiated, 42 were moderately differentiated and 12 were poorly differentiated. Five serum tumor markers of CA211, CA199, TPA, CA724 and NSE were selected optimally to create the optimal SVM model for diagnosis of oral squamous cell carcinoma. The accuracy, specificity, sensitivity and positive predictive value of the optimal SVM model were 88.54%, 93.13%, 84.05% and 92.57%, respectively.. From the results, SVM model combined with 5 optimal serum tumor markers is suggested to be used in the diagnosis of oral squamous cell carcinoma. Supported by Shanghai Leading Academic Discipline Project (Grant No.Y0203).

Topics: Adult; Aged; Aged, 80 and over; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Mouth Neoplasms; Phosphopyruvate Hydratase; Sensitivity and Specificity; Support Vector Machine

Lymphatic metastasis has always been regarded as a major prognostic indicator for disease progression and as a guide for therapeutic strategies to oral squamous cell carcinoma (OSCC), but to date, how tumor cells access and spread via the lymphatics have not been fully elucidated. Whether tumor cells metastasize by expansion and invasion of pre-existing peritumoral lymphatics or by the induction and invasion of newly formed lymphatics within tumors is controversial. In order to address this issue and find out the clinicopathological significance of intratumoral lymphangiogenesis, we investigated 86 archival specimens from patients with OSCC, quantitating lymph vessels by immunostaining with D2-40. We also quantified lymphatic invasion and examined the possible associations of all the above parameters with clinicopathological features and outcome. Higher intratumoral lymphatic density (ILD) and peritumoral lymphatic density (PLD) were both significantly associated with the presence of lymph node metastasis at the time of diagnosis and the outcome of the post-operation biopsy of 77 patients (P = 0.001). Higher ILD was significantly associated with a higher incidence of intratumoral lymphatic invasion, peritumoral lymphatic invasion and recurrence of tumor (P = 0.001 and P = 0.041 and P = 0.001, respectively). Patients with higher ILD exhibited shorter 5-year cumulative and disease-free survival (P = 0.001). Thus, lymphangiogenesis indeed occurs in oral squamous cell carcinoma; ILD might be used as an index to inflect the aggression of the disease, to evaluate the status of lymphatic metastasis, to separate patients at higher risk of an adverse clinical outcome.

Topics: Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease-Free Survival; Endothelium, Lymphatic; Female; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Keratins; Lymphangiogenesis; Lymphatic Metastasis; Male; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; Proportional Hazards Models

Oral submucous fibrosis (OSF) is characterized by abnormal collagen metabolism in the submucosal connective tissue. Its influence on the overlying epithelium is not known but about 14% of OSF cases undergo malignant transformation to squamous cell carcinoma indicating association with abnormality of the epithelium. Here, we have defined the keratin expression profile, by immunohistochemistry and quantitative image analysis, using a panel of 22 anti-keratin monoclonal antibodies on 28 OSF samples. We observed an increase of K1 and K10 in the suprabasal layers, induction of K6 in the basal layer and complete loss of K19 in the epithelium. Furthermore, there was increased K17 expression in the suprabasal layers, which correlated with disease severity. In a subset of the most severe OSF cases (14%), K17 expression was completely lost in the basal layer which might define them to be at most risk to undergo malignant transformation. There was no detectable expression of K8, K18, K7 and K9 and the expression of K4, K13, K14, K15 and K16 did not change in OSF. We propose that the altered keratin profiles could be useful as histological diagnostic markers and provide important insights into the pathogenesis of the disease and its predisposition to malignancy.

Topics: Biomarkers; Carcinoma, Squamous Cell; Case-Control Studies; Cell Transformation, Neoplastic; Gene Expression; Humans; Immunohistochemistry; Keratin-17; Keratinocytes; Keratins; Mouth Neoplasms; Oral Submucous Fibrosis; Phenotype; Photography, Dental; Precancerous Conditions; Severity of Illness Index

Topics: Diagnosis, Differential; Humans; Immunophenotyping; Keratins; Lymphatic Metastasis; Melanoma; Middle Aged; Mouth Neoplasms; Prognosis; Submandibular Gland Neoplasms

Oral squamous cell carcinoma (OSCC) is the most common malignant tumour in the oral and maxillofacial region, and has a poor prognosis. Cyfra 21-1 is a useful tumour marker for squamous cell carcinoma, but the clinical value of Cyfra 21-1 in OSCC has not been confirmed. In order to investigate the diagnostic and prognostic value of serum Cyfra 21-1 in primary OSCC patients, the preoperative serum Cyfra 21-1 concentration of 100 OSCC patients and 56 healthy subjects was detected by enzyme-linked immunosorbent assay (ELISA). The cut-off value was calculated with a receiver operating characteristic (ROC) curve, and prognostic analysis was performed using the Kaplan-Meier method and Cox regression models. The preoperative serum Cyfra 21-1 concentration in OSCC patients (1.18+/-1.20 microg/L) was significantly higher (t=6.585, P<0.001) than that in healthy subjects (0.40+/-0.16 microg/L). With a cut-off value of 0.65 microg/L, the diagnostic sensitivity and specificity was 0.570 and 0.964, respectively. There was significant correlation with tumour recurrence and survival rate: the higher the serum Cyfra 21-1; the higher the tumour recurrence rate and lower the survival rate. Serum Cyfra 21-1 was an independent prognostic factor for OSCC using univariate and multivariate Cox models.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Kaplan-Meier Estimate; Keratin-19; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Prognosis; Proportional Hazards Models; ROC Curve; Sensitivity and Specificity

Topics: Adenocarcinoma; Adult; Carcinoembryonic Antigen; Dermoid Cyst; Humans; Immunohistochemistry; Intestines; Keratins; Male; Mouth Floor; Mouth Neoplasms; Mucin-1

ATF3 is a highly conserved eukaryotic transcription factor that is ubiquitously upregulated transcriptionally during cellular responses to a variety of stresses, in particular DNA damage. However, the role of ATF3 in the DNA damage response is unclear. Transgenic mice that overexpress human ATF3 in basal epithelial cells under the control of the bovine keratin 5 (K5) promoter were constructed and characterized for epidermal alterations. Strong, nuclear expression of the exogenous ATF3 protein was seen in basal cells of the epidermis, hair follicles, and oral mucosa. Hyperplastic changes in the K5-expressing, outer root sheath (ORS) cells of the hair follicle were observed in young mice, resulting in multiple layers of ORS cells in the mature follicle and large aberrantly shaped follicles. Mild hyperplasia of the interfollicular epidermis was also noted, increasing with age. However, no epidermal tumors were identified in BK5.ATF3 mice observed for 16 mo. At 16 mo of age, most transgenic mice exhibited multi-focal areas of hyperplasia and dysplasia in the oral mucosa, with cellular atypia and underlying acute inflammatory changes. Neoplastic lesions were also seen in the oral cavity of BK5.ATF3 mice, including oral squamous cell carcinoma (60% incidence) and basal cell tumors with follicular differentiation (70% incidence), but not in non-transgenic FVB/N littermates. Heterogeneous nuclear expression (or stabilization) of p53 protein was seen in some oral dysplasias, with a patchy distribution primarily in the least differentiated layers of the lesions. This represents the first indication that ATF3 may have oncogenic properties in epithelial cells.

Topics: Activating Transcription Factor 3; Animals; DNA, Complementary; Gene Expression Regulation, Neoplastic; Genotype; Humans; Keratin-5; Keratins; Mice; Mice, Transgenic; Mouth Neoplasms; Plant Roots; Time Factors; Transcription Factors; Tumor Suppressor Protein p53

To investigate the clinicopathological characteristics and biologic behaviour of epithelioid hemangioendothelioma in the oral cavity.. The clinical features and pathological findings of nine cases with intraoral epithelioid hemangioendothelioma were reviewed, including immunohistochemistry study.. This series comprised seven males and two females aged 6-53 years (mean 28 years). The sites of the tumour included the tongue (n = 4), lip (n = 1), the gingiva and alveoli of the maxilla (n = 1), the gingiva and alveoli of the mandible (n = 1), buccal mucosa (n = 1), and the floor of the mouth (n = 1). A painless solitary mass was the most common presentation and was found in eight cases. On pathology, the tumour grew in short strands, cords or nests of polygonal to slightly spindled epithelioid cells in fibro-myxoid stroma, with formation of intracytoplasmic lumina. Tumour cells were immunoreactive to CD34, FVIIIRAg, and vimentin. Focal-positive cytokeration were observed in three cases. Immunoreactivity for S-100 protein, epithelial membrane antigen (EMA) and human herpesvirus (HHV)-8 was negative in all cases. Two cases recurred after surgical excision, but no patient developed local or distant metastasis.. Wide local excision with long-term follow-up seems to be the treatment of choice for intraoral epithelioid hemangioendothelioma because of their unpredictable biological behaviour and recurrence potential.

Topics: Adolescent; Adult; Antigens, CD34; Child; Female; Gingival Neoplasms; Hemangioendothelioma, Epithelioid; Herpesvirus 8, Human; Humans; Keratins; Lip Neoplasms; Male; Mandibular Neoplasms; Maxillary Neoplasms; Middle Aged; Mouth Floor; Mouth Mucosa; Mouth Neoplasms; Mucin-1; Neoplasm Recurrence, Local; Retrospective Studies; S100 Proteins; Tongue Neoplasms; Vimentin; von Willebrand Factor

Lymphatic metastatic characteristics of oral squamous cell carcinoma are not fully understood, for instance, skip metastasis is still controversial. The purposes of the present study was to explore the accuracy and applicability of immunohistochemical stain with cytokeratin combining semi-serial sections for detection of cervical lymph node metastasis of oral squamous cell carcinoma.. Regional lymph nodes (N=1638) were obtained from 26 patients with primary oral squamous cell carcinoma who underwent five level neck dissections. Semi-serial sections at an interval of 0.5mm was performed for each lymph node and cross-detected by immunohistochemical staining with cytokeratin and traditional hematoxylin-eosin staining (H-E) and their accuracies were compared.. Of 26 patients, 21 were detected having lymphatic metastasis by H-E staining and 26 by immunohistochemical detection; Of 1638 lymph nodes, 52 metastatic lymph nodes were detected by H-E staining while 162 by immunohistochemical detection. One case with cancer of the mouth floor being defined having skip metastasis was proved having no skip metastasis by the immunohistochemical detection.. The immunohistochemical detection method with semi-serial sections has higher accuracy than the traditional H-E staining and its application may present a need to re-evaluate the neck metastatic patterns of oral squamous cell carcinoma.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Eosine Yellowish-(YS); Female; Hematoxylin; Humans; Immunoenzyme Techniques; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Mouth Floor; Mouth Neoplasms; Neck; Neoplasm Staging; Sensitivity and Specificity; Staining and Labeling

Saliva is the body fluid in the oral cavity and contacts directly with oral mucosa. As a detective media, it is acceptable and non-traumatic. Cyfra 21-1, being the soluble fragment of cytokeratin 19(CK19), correlates well with oral squamous cell carcinoma (OSCC).. To investigate the saliva Cyfra 21-1 concentrations in OSCC patients and healthy persons, and the correlation between the Cyfra 21-1 concentration in saliva and the CK19 expression in tissue from OSCC patients.. Saliva Cyfra 21-1 concentration was detected by ELISA in 30 OSCC patients and 30 healthy persons; CK19 protein expression and CK19 mRNA level were, respectively, detected by immunohistochemistry and fluorescent real-time RT-PCR in cancerous and paracancerous tissues from 33 OSCC patients.. Saliva Cyfra 21-1 concentration in OSCC patients (85.95+/-78.00 microg/L) was significantly higher than that in healthy persons (42.27+/-40.84 microg/L) (P=0.009); it was also significantly higher in the patients suffering later tumour recurrence (130.95+/-66.38 microg/L) than that in the patients without tumour recurrence (74.84+/-63.45 microg/L) (P=0.023). CK19 protein expression increased significantly in OSCC tissues (P<0.001) with positive rate of 90.9%, CK19 mRNA level in cancerous tissues was 2.21 folds higher than that in paracancerous tissues (P=0.020); significant correlation was found between tissue CK19 protein expression and tissue CK19 mRNA level (P=0.003), and great correlation was found between tissue CK19 protein expression and saliva Cyfra 21-1 concentration (P=0.051).. The increased CK19 expression in OSCC tissues plays an important role in the increase of saliva Cyfra 21-1 concentration. Potential clinical value of saliva Cyfra 21-1 detection is suggested for OSCC. Further studies are encouraged to reveal the real diagnostic and prognostic value of detecting saliva Cyfra 21-1 concentration for OSCC.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunohistochemistry; Keratin-19; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Precancerous Conditions; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; ROC Curve; Saliva; Statistics, Nonparametric; Survival Rate

Extended histopathologic work-up has increased the detection of micrometastasis in sentinel lymph nodes in malignant melanoma and breast cancer. The aim of this study was to examine if (A) step-sectioning of the central 1000 microM at 250 microM levels with immunostaining were accurate when compared with (B) step-sectioning and immunostaining of the entire sentinel lymph node at 250 microM levels.. Forty patients with T1/T2 cN0 oral cancer were enrolled. Three patients were excluded. In one patient no sentinel lymph node was identified. The remaining two had unidentified sentinel lymph nodes due to lymphoscintigraphic and surgical sampling error. The central 1000 microM of 147 sentinel lymph nodes were step-sectioned in 250-microm intervals and stained with hematoxylin and eosin and CK-KL1. All lymph nodes were recorded as negative or positive for macrometastases or micrometastases. After inclusion of the last patient the residual tissue of the lymph nodes was totally step-sectioned at 250-microm intervals and re-classified. The tumor deposits were divided into macrometastases and micrometastases and ITC.. Method (A) upstaged 17 lymph nodes and 11 patients compared with method (B), which upstaged 22 lymph nodes and 11 patients. Seven of the patients with positive lymph nodes did not change stage. However, four lymph nodes changed from micrometastases to macrometastases. One patient changed from a micrometastasis to four micrometastases. One pN2c patient with bilateral micrometastases did not change stage, but an additional ipsilateral lymph node with a micrometastasis was identified.. Larger tumor deposits and more metastases are identified by more extensive sectioning of the sentinel lymph nodes. None of the patients was false-negative due to histopathologic sampling error, but the results indicate that central step-sectioning of the central 1000 microM cannot completely be relied upon for accurate staging of the patients.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Coloring Agents; Eosine Yellowish-(YS); Female; Fluorescent Dyes; Follow-Up Studies; Hematoxylin; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Microtomy; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Sentinel Lymph Node Biopsy; Survival Rate

Intermediate filaments are involved in cell migration and intracellular signal transduction pathways. In a variety of organs, the expression of distinct intermediary filaments are further associated with distinct steps of malignant transformation. In this study, we seeked to define the cytokeratin (Ck) expression pattern in oral leukoplakia and oral squamous cell carcinoma (OSCC). One hundred and ninety-two patients with OSCC, 117 patients with oral leukoplakia without dysplasia (OL) and 23 with oral leukoplakia with dysplasia (squamous intraepithelial neoplasia) (OLD) of the oral cavity were investigated for the immunohistochemical expression of Ck 5-6, Ck 8/18, Ck 1 Ck 10, Ck 14, Ck 19 using the tissue microarray technique. Correlations between clinical features and the expression of cytokeratins were evaluated statistically by chi2 tests. The expression of Ck 8/18, Ck 19 and Ck 1 was seen in 3.1% (Ck 8/18), 12.5% (Ck 19), 75.4% (Ck 1) of all leukoplakias, 1.0% (Ck 8/18), 9.4% (Ck 19), 76.8% (Ck 1) in OL, 13.0% (Ck 8/18), 27.3% (Ck 19), 68.4% (Ck 1) in OLD and was significantly associated with the degree of dysplasia (Ck 8/18 p<0.01; Ck 19 p<0.01; Ck 1 p<0.01) and the acquisition of invasive growth properties. The highest frequencies were observed in invasive squamous cell carcinomas. The expression of Ck 8/18 and Ck 19 in transformed oral lesions can be regarded as an early feature in the pathogenesis of invasive OSCC. However, the aberrant expression of Ck 8/18 and Ck 19 in an even higher frequency in invasive carcinomas characterizes the expression of typical glandular cytokeratins as a general progression marker in squamous cell carcinomas. These results can be interpreted as first hints that oral leukoplakias with an expression of Ck 8/18 or 19 independent of dysplasia, should be resected totally since they might indicate an increased progression potential.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Female; Humans; Keratins; Leukoplakia; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging

Oral submucous fibrosis (OSF) is a pre-malignant condition caused by habitual use of areca nut, affecting the oro-pharynx and characterized by progressive fibrosis. Alteration of cytokeratin (CK) expression has been documented in leukoplakia and oral cancer (OC). However, very little is known of CK alterations in OSF. The present study was carried out to characterize the CK profile in OSF and ascertain if this could be used as a surrogate marker for malignant transformation.. Paraffin-embedded tissues of OSF (n = 50), normal (n = 10) and OC (n = 10) were stained with pancytokeratin (PanCK), high molecular weight cytokeratin (HMWCK), CKs 18, 14, 8, 5, 4 and 1 by immunohistochemistry.. Significant difference in the CK staining pattern was seen between normal, OSF and cancer. Significant changes in OSF included increased intensity of staining for PanCK and HMWCK, aberrant expression of CK8 and decreased expression of CKs 5 and 14.. Cytokeratin profile of OSF was significantly different from normals for PanCK, HMWCK, CK8, 5 and 14 suggesting their potential to be used as surrogate markers of malignant transformation. Further studies will help in better defining the nature and clinical implications of these alterations.

Topics: Biomarkers, Tumor; Cell Transformation, Neoplastic; Coloring Agents; Female; Humans; Immunohistochemistry; Keratins; Male; Molecular Weight; Mouth Neoplasms; Oral Submucous Fibrosis; Precancerous Conditions

Intermediary filaments are involved in cell motility and cancer progression. In a variety of organs, the expression of distinct intermediary filaments are associated with patient prognosis. In this study, we seeked to define the prognostic potential of cytokeratin and vimentin expression patterns in squamous cell carcinomas (SCC's) of the oral cavity.. 308 patients with histologically proven and surgically treated squamous cell carcinomas of the oral cavity were investigated for the immunohistochemical expression of a variety of intermediary filaments including high- and low-molecular weight cytokeratins (Ck's), such as Ck 5/6, Ck 8/18, Ck 1, CK 10, Ck 14, Ck 19 and vimentin, using the tissue microarray technique. Correlations between clinical features and the expression of Cytokeratins and vimentin were evaluated statistically by Kaplan-Meier curves and multivariate Cox regression analysis.. The expression of Ck 8/18 and Ck 19 were overall significantly correlated with a poor clinical prognosis (Ck 8/18 p = 0.04; Ck19 p < 0.01). These findings could also be reproduced for Ck 8/18 in primary nodal-negative SCC's and held true in multivariate-analysis. No significant correlation with patient prognosis could be found for the expression of the other cytokeratins and for vimentin.. The expression of Ck 8/18 in SCC's of the oral cavity is an independent prognostic marker and indicates a decreased overall and progression free survival. These results provide an extended knowledge about the role of intermediary filament expression patterns in SCC's.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Gene Expression Profiling; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Prognosis; Survival Analysis

To investigate the incidence of central nodal necrosis (CNN) in the cervical lymph nodes of patients with oral squamous cell carcinoma (SCC) and the factors that influence the formation of CNN.. Lymph nodes shown as CNN on computed tomography (CT) films in 107 lymph nodes from 27 patients with oral SCC were selected. Lymph nodes with CNN on CT films were compared with the pathological findings of lymph nodes on specimens. We compared many kinds of factors influencing the formation of CNN, including the differentiated type, with the incidence of CNN.. Significant relationships were found between the incidence of CNN in metastatic lymph nodes and the presence of well-differentiated SCC and the presence of keratinization in tumour cells.. The results indicated that if a patient had SCC with low-grade differentiation, CNN in small lymph nodes would be difficult to detect on CT scan. Therefore, noting changes in lymph node density in the absence of CNN on CT scans is necessary in case the primary tumour is low-grade SCC.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neck; Neck Dissection; Necrosis; Neoplasm Staging; Tomography, X-Ray Computed

Adenoid squamous cell carcinoma (ASCC) is an uncommon but well-recognized variant of squamous cell carcinoma that was first described by Lever in 1947. ASCC has been reported to originate in the sun-exposed skin of the head and neck and in other sites. An additional case of ASCC is reported here. The patient was a 64-year-old Japanese woman who requested examination of a reddish lesion on the left floor of the mouth. The biopsy material was diagnosed as squamous cell carcinoma. Clinical examination showed a well-circumscribed, 20 x 10 mm-sized lesion, which was categorized as cT2cN0cm 0. Tumor resection was therefore performed. Histologically, most parts of the lesion were conventional squamous cell carcinoma in situ, but the invasive part consisted of ASCC with gland-like or reticular appearance. The latter part was negative for mucin staining. Immunohistochemically, this lesion was positive for pancytokeratin, high-molecular-weight keratin, cytokeratin (CK) 7/8, CK19, E-cadherin and p53, but negative for vimentin, CK20, and S-100 protein. The Ki-67 labeling index was 50.3% in the ASCC part and 34.5% in the carcinoma in situ part. These findings and a review of the literature indicate that a gland-like feature of ASCC is associated with the loss of cell adhesion in the center of the cancer nests, and it can be confirmed simply by mucin staining to be neither an adenosquamous carcinoma nor ductal involvement of conventional squamous cell carcinoma.

Topics: Cadherins; Carcinoma, Adenosquamous; Carcinoma, Mucoepidermoid; Carcinoma, Squamous Cell; Cell Adhesion; Cell Proliferation; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Middle Aged; Mouth Neoplasms; Mucins; Tumor Suppressor Protein p53

The p16 and retinoblastoma (Rb) gene products are part of the retinoblastoma pathway controlling the G1-S transition of the cell cycle. Few studies on the expression of p16 and retinoblastoma proteins in oral cavity squamous carcinomas have been conducted.. To correlate the expression of p16 and retinoblastoma proteins to clinicopathological characteristics in these tumours.. 45 patients with resected oral cavity squamous carcinoma were selected, for whom this was the initial treatment and who were followed up for 5 years or until death. Immunohistochemical stains with antibodies to the Rb and p16 gene products were carried out on paraffin wax-embedded tissue. Data on clinicopathological features such as tumour differentiation, nodal status, stage and survival outcome were collected.. Retinoblastoma expression was seen in 39 of 45 (87%) patients and p16 expression in 6 of 45 (13%) patients. A significant inverse correlation was observed between retinoblastoma and p16 expression as nearly all retinoblastoma negative cases were p16 positive, and vice versa. When examined for clinicopathological correlates, it was found that all 39 tumours that expressed retinoblastoma displayed marked keratinisation and were of low-moderate histological grade. Conversely, five of the six tumours that expressed p16 were found to be poorly differentiated, with minimal keratin expression.. Salient relationships were seen between expression of retinoblastoma and p16 and keratinisation. A marked loss of keratin production was evident in the tumours that expressed p16. Tumours expressing retinoblastoma were seen to exhibit more widespread keratinisation. In addition, an inverse staining pattern was found for retinoblastoma and p16 as retinoblastoma-expressing tumours were nearly universally p16 negative and vice versa. No correlation of expression of either p16 or retinoblastoma was found with survival or stage. A link between the histologically observable morphology and expression of cell cycle regulatory protein with the expression of p16 and retinoblastoma has been suggested with keratinisation and differentiation of status.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Cyclin-Dependent Kinase Inhibitor p16; Female; Follow-Up Studies; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Proteins; Neoplasm Staging; Retinoblastoma Protein; Survival Analysis

The main goal of sentinel lymph node (SLN) detection in head and neck squamous cell carcinomas is to limit neck dissections to pN+ cases only. However, intraoperative + diagnosis cannot be routinely done using the current gold standard, serial step sectioning with immunohistochemistry. Real-time quantitative reverse transcription-PCR (RT-PCR) is potentially compatible with intraoperative use, proving highly sensitive in detecting molecular markers. This study postoperatively assessed the accuracy of quantitative RT-PCR in staging patients from their SLN.. A combined analysis on the same SLN by serial step sectioning with immunohistochemistry and quantitative RT-PCR targeting cytokeratins 5, 14, and 17 was done in 18 consecutive patients with oral or oropharyngeal squamous cell carcinoma and 10 control subjects.. From 71 lymph nodes examined, mRNA levels (KRT) were linked to metastasis size for the three cytokeratins studied (Pearson correlation coefficient, r = 0.89, 0.73, and 0.77 for KRT 5, 14, and 17 respectively; P < 0.05). Histopathology-positive SLNs (macro- and micrometastases) showed higher mRNA values than negative SLNs for KRT 17 (P < 10(-4)) and KRT 14 (P < 10(-2)). KRT 5 showed nonsignificant results. KRT 17 seemed to be the most accurate marker for the diagnosis of micrometastases of a size >450 mum. Smaller micrometastases and isolated tumor cells did not provide results above the background level. Receiver operating characteristic curve analysis for KRT 17 identified a cutoff value where patient staging reached 100% specificity and sensitivity for macro- and micrometastases.. Quantitative RT-PCR for SLN staging in cN(0) patients with oral and oropharyngeal squamous cell carcinoma seems to be a promising approach.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Keratin-14; Keratin-5; Keratins; Lymph Nodes; Lymphatic Metastasis; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Oropharyngeal Neoplasms; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Sentinel Lymph Node Biopsy

Adequate brushing of oral mucosa is important for accurate human papillomavirus (HPV) detection in potentially malignant (oral leukoplakia [OL], oral lichen planus [OLP]) and malignant (oral squamous cell carcinoma [OSCC]) lesions. Since various factors may limit the adequacy of oral brushing and, consequently, the accuracy of HPV detection, modified sampling procedures should be evaluated for their effect on HPV frequency and/or types detected.. To compare the HPV frequency in samples obtained by brushing the lesion site with the frequency in samples obtained by brushing an apparently normal adjacent site. The correlation between HPV frequency and keratinization of the site affected by the lesion, as well as sociodemographic variables (age, sex, smoking and drinking habits), was also examined.. HPV DNA was detected in brushing samples from 50 patients with OL, 49 with OLP, and 17 with OSCC. Polymerase chain reaction (PCR) amplification was performed by MY09/MY11 and GP05+/GP06+ primers; the HPV type was identified by DNA sequencing and a reverse hybridization (line probe) assay. Data were analyzed by the Z test, the Fisher's exact test, the chi-square test, odds ratio (OR), and a logistic regression model.. HPV DNA was detected in 22% of samples from lesion sites and in 16% of samples from adjacent sites (p = 0.22) in patients with OL, in 24.5% and 22.4% of samples from lesion and adjacent sites, respectively, in patients with OLP (p = 0.40), and in 35.3% and 41.2% of samples from lesion and adjacent sites, respectively, in patients with OSCC (p = 0.36). Lesions adjacent to HPV-positive normal sites had an increased rate of HPV detection (OR = 30; 95% CI 9.57, 94.1). HPV-18 was the most frequent genotype, followed by HPV-6, -16, -33, and -53. HPV prevalence was reduced in lesions at keratinized sites (14.5%) compared with non-keratinized sites (34.4%; p = 0.007; OR = 0.32; 95% CI 0.13, 0.81).. In patients with OL, OLP, or OSCC, a high prevalence of HPV infection was shown in apparently normal sites adjacent to lesion sites infected by HPV. The lower HPV frequency in lesions at keratinized sites suggests that HPV detection by lesion brushing is affected by keratinization. The keratinized epithelium may be less susceptible to HPV infection or, alternatively, the highly proliferative activity in non-keratinized sites may predispose to HPV infection.. Results from this study indicate that taking samples from normal sites adjacent to oral lesions may be of value in HPV detection, particularly when the lesions are located at keratinized sites. This sampling procedure may allow more accurate diagnosis of HPV infection compared with sampling only the lesion site, and may also represent a reliable method to investigate the biological characteristics of HPV infection and related oral carcinogenesis.

Topics: Alphapapillomavirus; Carcinoma, Squamous Cell; DNA, Viral; Female; Humans; Keratins; Lichen Planus, Oral; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Papillomavirus Infections

In this study we evaluated pedunculated oncocytic carcinoma (OC) in the buccal mucosa via immunohistochemical and ultrastructural studies.. An 84-year-old man was referred to our clinic with a pedunculated mass about 4 cm in diameter in the right buccal mucosa. An incision biopsy revealed the diagnosis of oncocytic tumor, and enucleation was performed. The tumor was stained for immunohistochemical analysis using the ABC method and antibodies against cytokeratin (CK), epithelial membrane antigen (EMA), desmin, S-100 protein and muscle-specific actin, respectively. The tumor was stained with uranyl acetate and lead citrate for visualization by electron microscopy.. Histopathology results revealed that the tumor consisted of oncocytic cells, characterized by eosinophilic and granular cytoplasm, and atypical nuclei. These cells had infiltrated local blood vessels, salivary glands and muscles. Immunohistochemical staining indicated that these cells were positive for CK and EMA, and negative for desmin, muscle-specific actin and S-100 protein. Electron microscopy revealed numerous dilated cytoplasmic mitochondria, and the cell contours and nucleic shapes of tumor cells were often irregular.. Because the histopathologic features of OC are similar to those of benign oncocytoma, the diagnosis of OC must be confirmed by a combination of clinical and ultrastructural characteristics.

Topics: Adenocarcinoma; Aged, 80 and over; Cheek; Humans; Immunohistochemistry; Keratins; Male; Microscopy, Electron, Transmission; Mitochondria; Mouth Mucosa; Mouth Neoplasms; Mucin-1; Neoplasm Invasiveness

Oral squamous cell carcinoma develops through a multistep of genetic mutations, and the process can be morphologically recognized as oral epithelial dysplasia. To evaluate the hypothesis that distributional alterations of proliferating and stem cells may be a useful index to estimate the grading and development of epithelial dysplasia, we examined the distribution patterns according to stratified cell layers.. Sixty-two oral dysplasia cases according to the histological grades were immunohistologically examined and the nuclear expression of Ki-67 and p63 antigens was counted according to epithelial layers as labeling index.. The Ki-67 labeling index in the basal and suprabasal layers and that of p63 in the basal layer showed a significant difference between low- and high-grade groups of epithelial dysplasia.. The architectural alteration of proliferating cell and stem cell distribution in the layers of epithelial dysplasias may provide useful information to evaluate the grading of oral epithelial dysplasias.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Cell Proliferation; Epithelium; Female; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Male; Membrane Proteins; Middle Aged; Mouth Neoplasms; Neoplasm Proteins; Neoplastic Stem Cells; Precancerous Conditions; Tumor Suppressor Protein p53

A 10-year-old, female, pot-bellied pig (Sus scrofa) experienced a 3-month history of reduced appetite, dysphagia, and weight loss. Clinical examination revealed a mass in the left part of the oral cavity extending from the hard to the soft palate. At necropsy, a firm, white, poorly demarcated ulcerated mass at the left hard and soft palate with metastases to the left retropharyngeal lymph node and the lung was observed. Additional findings included a uterine adenocarcinoma, a hepatocellular adenoma, and nodular hyperplasias in spleen and adrenal glands. Histologically, the poorly demarcated, infiltrative growing oral mass consisted of islands, cords, and single epithelial cells with moderate squamous differentiation. Cells were strongly positive for cytokeratin by immunohistochemistry. Similar cells were found in the left retropharyngeal lymph node and the lung. The present findings represent the first report of a metastasizing oral squamous cell carcinoma in a pig.

Topics: Animals; Carcinoma, Squamous Cell; Fatal Outcome; Female; Immunohistochemistry; Keratins; Mouth Neoplasms; Neoplasm Metastasis; Sus scrofa; Swine; Swine Diseases

To analyze the lymphatic distribution of metastatic carcinomatous cells in cervical lymph nodes in head and neck squamous cell carcinoma (HNSCC).. We retrospectively reviewed 119 patients treated in our hospital for HNSCC (1999-2004). Topography of the neck dissection specimens was prospectively classified according to the classification of Robbins. The 4000 lymph nodes were analyzed by optical microscopy using hematoxylin-eosin-safran (HES) staining. In cases of negative results in level II, cytokeratin (AE1/AE3) immunodetection was performed.. Metastases were visualized using HES in 6.4% of lymph nodes for oral cavity, and 4.7% of oropharyngeal, 4.4% of hypopharyngeal, and 1.3% of endolaryngeal cancers. The highest incidence of nodal metastasis was observed in level IIa (P < 0.01). In eight patients (6.7%) with lymph node metastases, level II was spared. In these patients, all 134 nodes histologically negative on HES were confirmed to be negative by IHC.. Level IIa is the main level involved in regional metastases of HNSCC, regardless of the primary site of cancer. However, in eight (6.7%) patients, level II was spared, as confirmed by IHC. In these cases, level II did not represent the first step of drainage from the tumor. The sentinel lymph node technique in HNSCC is discussed in light of these results.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Coloring Agents; Female; Fluorescent Dyes; Head and Neck Neoplasms; Humans; Keratins; Laryngeal Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neck; Neck Dissection; Neoplasm Staging; Pharyngeal Neoplasms; Prospective Studies; Retrospective Studies; Sentinel Lymph Node Biopsy

TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) preferentially induces apoptosis of cancer cells without toxicity in normal cells. TRAIL plays an important role in host immune surveillance against tumor metastasis. Cathepsin B (CB) is a mediator of apoptosis whose activity is regulated by its inhibitors, known as cystatins. We examined the TRAIL-mediated cytotoxicity rates of clonally-related primary and metastatic oral cancer (OC) cells and correlated them with the expression levels of TRAIL receptors, cathepsin B and cystatins A, B, C and M. Two pairs of primary (686Tu and 101A) and metastatic (686Ln and 101B) OC cell lines were treated with various concentrations (5 to 1000 ng/ml) of recombinant human TRAIL protein for 14 h, and cell viability and apoptotic rate were determined. In both pairs of cell lines, primary OC cells revealed greater susceptibility to TRAIL than their metastatic counterparts. The protein synthesis inhibitor cycloheximide markedly increased the TRAIL sensitivity of these cell lines, whereas the CB-specific chemical inhibitor CA-074 markedly reduced the sensitivity of primary OC cells to TRAIL. DNA laddering and M30 CytoDEATH immunodetection assays confirmed that TRAIL-induced OC cell death is an apoptotic process. Expression levels of TRAIL death (DR4 and DR5) and decoy (DcR1 and DcR2) receptors were not different between primary and metastatic OC cells. However, expression levels of cystatins were higher in metastatic OC cells than in their respective primary cells, whereas CB levels remain unchanged. Cathepsin B is a mediator of TRAIL-induced apoptosis in OC cells. Elevated levels of cystatins in metastatic OC cells may cause their greater resistance to TRAIL-induced apoptosis. Our data suggest that high expression of cystatins in OC cells may confer a metastatic phenotype by enhancing their resistance to TRAIL.

Topics: Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; Caspase 3; Caspase 8; Caspases; Cathepsin B; Cell Survival; Cystatins; Dipeptides; Drug Resistance, Neoplasm; Humans; Keratins; Lymphatic Metastasis; Male; Membrane Glycoproteins; Mouth Neoplasms; Receptors, Tumor Necrosis Factor; Recombinant Proteins; TNF-Related Apoptosis-Inducing Ligand; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

We investigated the relationship between cell growth and differentiation and COX-2 expression in oral squamous cell carcinoma (SCC) in vitro and in vivo. Treatment of SCC25 oral squamous carcinoma cells with sodium butyrate (SB) at 0.5-5 mM or all-trans retinoic acid (ATRA) at 3-300 microM inhibited cell growth and induced apoptosis in a dose-dependent manner with concomittant increases in expression of keratin 13, p21WAF1/Cip1 and p27Kip1 and decreases in expression of COX-2. These effects were more pronounced with SB than with ATRA. Injection of SB or ATRA near SCC25-derived tumors in nude mice resulted in inhibition of growth and elevation of differentiation of the tumor accompanied by marked keratinization and increased expression of keratin 13 and decreased expression of COX-2. These results show that the differentiation-inducing agents, particularly SB, suppress growth of oral squamous carcinoma cells through apoptosis and induce cell differentiation possibly through mechanisms involving COX-2, p27Kip1 and/or p21WAF1/Cip1 in vitro and in vivo.

Topics: Animals; Apoptosis; Blotting, Western; Butyrates; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Cyclooxygenase 2; DNA Primers; Female; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Isobutyrates; Keratins; Membrane Proteins; Mice; Mice, Inbred BALB C; Mice, Nude; Mouth Neoplasms; Prostaglandin-Endoperoxide Synthases; Reverse Transcriptase Polymerase Chain Reaction; RNA; Time Factors; Tretinoin; Tumor Suppressor Proteins

This study characterizes the morphologic features and the endogenous fluorescence in the stratified squamous epithelia of the 7,12-dimethylbenz(a)anthracene-treated hamster cheek pouch model of carcinogenesis using multiphoton laser scanning microscopy (MPLSM). MPLSM allows high-resolution, three-dimensional image data to be collected deeper within thick tissue samples with reduced phototoxicity compared with single-photon imaging. Three-dimensional image stacks of normal (n = 13), precancerous (dysplasia, n = 12; carcinoma in situ, n = 9) and cancerous tissue [nonpapillary squamous cell carcinoma (SCC), n = 10, and papillary SCC, n = 7] sites in the hamster cheek pouch were collected in viable, unsectioned tissue biopsies at a two-photon excitation wavelength of 780 nm. Five features were quantified from the MPLSM images. These included nuclear density versus depth, keratin layer thickness, epithelial thickness, and the fluorescence per voxel in the keratin and epithelial layers. Statistically significant differences in all five features were found between normal and both precancerous and cancerous tissues. The only exception to this was a lack of statistically significant differences in the keratin fluorescence between normal tissues and papillary SCCs. Statistically significant differences were also observed in the epithelial thickness of dysplasia and carcinoma in situ, and in the keratin layer thickness of dysplasia and SCCs (both nonpapillary and papillary). This work clearly shows that three-dimensional images from MPLSM of endogenous tissue fluorescence can effectively distinguish between normal, precancerous, and cancerous epithelial tissues. This study provides the groundwork for further exploration into the application of multiphoton fluorescence endoscopy in a clinical setting.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinogens; Carcinoma in Situ; Carcinoma, Squamous Cell; Cheek; Cricetinae; Epithelial Cells; Fluorescence; Keratins; Male; Mesocricetus; Microscopy, Confocal; Microscopy, Fluorescence, Multiphoton; Mouth Neoplasms; Precancerous Conditions

Autofluorescence spectroscopy and Raman spectroscopy have been suggested for lesion diagnostics. We investigate the information contained in autofluorescence and Raman spectra recorded from oral tissue slices of various lesion types. Thirty-seven human oral mucosa lesions were biopsied and freeze-dried. Complete autofluorescence images and spectra were recorded from 20 microm sections. Raman spectra were acquired from the same positions for 12 of the sections. Cluster analysis was applied to find any relationship between spectral shape and lesion type or cell layer. Autofluorescence images showed high intensities for keratin layers and connective tissue, but hardly any for the epithelium. Autofluorescence spectra were centered around 520 nm and did not show specific spectral features. No clustering with regard to lesion type or cell layer was observed. Raman spectra allowed for reliable classification into cell layers, but differences between lesion types were not significant in this study. Autofluorescence spectra of freeze-dried oral mucosa sections did not contain useful information. A more comprehensive study is required for Raman spectra.

Topics: Carcinoma, Squamous Cell; Cluster Analysis; Connective Tissue; Epithelium; Freeze Drying; Humans; Keratins; Mouth Mucosa; Mouth Neoplasms; Spectrometry, Fluorescence; Spectrum Analysis, Raman

To investigate clinicopathological and immunohistochemical features of 31 cases of oral teratoid cyst.. Thirty-one cases of oral teratoid cyst according with Meyer's diagnosis criteria were retrospectively studied and their histochemistry and immunohistochemistry were performed from the files of Department of Pathology, School of Stomatology, Wuhan University between 1963 and 2003.. Twenty-seven cases (87.10%) were children, and twenty-four cases were congenital. The ratio of male-to-female was 1:0.55, and the affected sites were floor of mouth (22 cases) and tongue (8 cases). Clinical findings were nonspecific, and prognosis was good following complete excision. Histology indicated that squamous, respiratory and/or gastrointestinal epithelium consistiuted basic structure of teratoid cyst in addition to simple cuboidal epithelium in 8 cases. Antibody against AE1/AE3 was strongly expressed and CK16 was weak in four types of epithelial lining of oral teratoid cyst. Expression of AE1, CK7, 8/18, 19 varied in superficial, suprabasal and basal cells of squamous epithelium but were strong in respiratory, gastrointestinal and simple columnar epithelium; only gastrointestinal epithelium expressed CK20 heterogeneously.. Oral teratoid cyst showed the highest incidence in children, and floor of mouth and tongue were mostly affected sites. Features of histology were complex, and immunohistochemistry indicated that epithelium of oral teratoid cyst shared similar patterns of cytokeratin with counterpart of normal tissues, showing different origin and differentiation of epithelial lining of the present cyst.

Topics: Adolescent; Adult; Child; Child, Preschool; Female; Humans; Immunohistochemistry; Infant; Keratins; Male; Mouth Neoplasms; Retrospective Studies; Teratoma; Young Adult

This study aimed to investigate the role of underlying fibroblasts on morphogenesis of in vitro epithelium reconstituted with normal and neoplastic human oral keratinocytes at various stages of malignant transformation. Primary normal human oral keratinocytes (NOKs), early neoplastic/dysplastic human oral keratinocytes (DOK cell line), and neoplastic human oral keratinocytes (PE/CA-PJ 15 cell line) were organotypically grown on top of a collagen type I matrix with or without primary normal human oral fibroblasts. Morphogenesis of the reconstituted epithelia was assessed by histomorphometry, immunohistochemistry (Ki-67, cyclin D1, cytokeratin 13 (CK13), collagen IV, E-cadherin, p53, CD40), and the terminal deoxynucleotidyl transferase-mediated dUTP in situ nick end-labelling method. Reproducible in vitro models of multistage oral carcinogenesis were established. Presence of fibroblasts in the collagen matrix significantly increased cell proliferation in all three models (p<0.05), and induced an invasive pattern of growth in the neoplastic cell lines (p<0.05). In normal, but not in neoplastic oral keratinocytes fibroblasts induced the expression of CD40, and polarized the expression of E-cadherin and p53 to the basal cell layer. In both normal and early neoplastic keratinocytes (DOK cell line), fibroblasts induced the expression of CK13 and collagen IV. In the neoplastic oral keratinocytes (PE/CA-PJ 15 cell line), the presence of underlying fibroblasts did not change the expression of any of the protein markers assessed. This study showed that (1) major steps of oral carcinogenesis can be reproduced in vitro, and (2) the tight control exerted by fibroblasts on epithelial morphogenesis of in vitro reconstituted normal human oral mucosa is gradually lost during neoplastic progression.

Topics: Biomarkers; Cadherins; Carcinoma; Cell Differentiation; Cell Transformation, Neoplastic; Collagen Type IV; Epithelium; Fibroblasts; Humans; Keratinocytes; Keratins; Morphogenesis; Mouth Mucosa; Mouth Neoplasms

Squamous cell carcinoma (SCC) is the most common malignant head and neck tumor and is responsible for more than 90% of head and neck cancers and accounts for 4.5% of all malignant tumors in males and 3.5% in females in South Korea. The purpose of this study was to investigate the correlation of suggested clinico-pathological prognostic factors such as gender, age, T score (T number in TNM), clinical stage, proliferation, invasion index, and lymph node metastasis to the survival of SCC patients in Korea. Furthermore, cytokeratin (CK), carcinoembryonic antigen (CEA), and recently documented apoptosis related protein, survivin, were analyzed by RT-PCR. In 113 patients, survival curves were estimated by the Kaplan-Meier method and nominal or numeric variable influence on survival was studied by Univariate and Multivariate Regression analysis (Cox proportional hazards model). Univariate analysis demonstrated that gender and age factor had no significant effect on survival rate. T score, on the other hand, significantly influenced survival and univariate analysis demonstrated that Stage 4 group had a significantly lower survival rate than the other stage groups but differentiation and invasion index factors had no significant effect on survival rate. Using a 50% cut-off point, patients with lower PCNA scores showed no survival advantages over those with higher PCNA scores but lymph node metastasis was a significant survival predictor in univariate analysis. In addition, lymph node CK and survivin mRNA expression have significant effects on OSCC patient survival rate. This means that prognostic value can be amplified by coincident analysis of T score, pathologically confirmed lymph node metastasis, and lymph node CK or survivin mRNA expression. Multivariate analysis using Cox's proportional hazards model, clinical TNM stage and lymph node survivin mRNA expression were independent OSCC prognostic factors, which support cancer staging based on the TNM as a powerful prognostic variable and lymph node survivin expression might provide predictive information for OSCC patient survival.

Topics: Adolescent; Adult; Age Factors; Aged; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Cell Differentiation; Female; Humans; Inhibitor of Apoptosis Proteins; Keratins; Lymphatic Metastasis; Male; Microtubule-Associated Proteins; Middle Aged; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Proteins; Neoplasm Staging; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sex Factors; Survival Analysis; Survivin

To identify differentially expressed genes during the development of oral malignancy, differential display, northern blotting, reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical analyses were undertaken using oral squamous cell carcinoma (OSCC) and leukoplakia tissues. Significantly higher levels of keratin (Ker)-14 and -17 mRNAs, combined with lower levels of Ker-4, Ker-13 and transglutaminase 3 (TG-3) transcripts, were observed in OSCC and severely dysplastic tissues, whereas this expression profile was reversed in hyperplasia and in mild to moderate dysplasia. The expression of Ker-4 and Ker-13 was elevated in density-arrested OSCC cell lines (Ca9-22, HSC-2, -3 and -4) but the expression of Ker-17 mRNA was elevated in these cells, regardless of the growth conditions. In addition, Ker-4 and Ker-13 proteins were predominantly expressed in moderate dysplasia and hyperplasia, whereas Ker-17 was markedly expressed in OSCC tissues. The expression patterns of these genes could therefore be an important determinant of the manifestation of oral malignancy.

Topics: Adult; Aged; Aged, 80 and over; Calcium-Binding Proteins; Carcinoma, Squamous Cell; Disease Progression; Female; Gene Expression Regulation, Neoplastic; Humans; Hyperplasia; Immunoenzyme Techniques; Keratin-14; Keratins; Leukoplakia, Oral; Male; Middle Aged; Mouth Neoplasms; Neoplasm Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Transglutaminases; Tumor Cells, Cultured

The aim of this study was immunolabeling oncoproteins Ck14, p53, p21 and Bcl-2 in order to evaluate their expression in premalignant and malignant stomatological lesions in oral epithelial, and to compare this expression with exfoliative cytology alterations in the same patients. It was studied biopsies and cytologies of 13 subjects with oral lichen planus, with or without Human Papilloma Virus (HPV), leukoplakia and squamous cell carcinoma clinically diagnosed and confirmed by anatomopathological studies. The oral lichen planus lesion presented binuclei orange cells; and in leukoplakia lesions only orange stained was observed; meanwhile koilocytes, inflammatory cells, enlarge nuclear volume and pathogenic microorganisms were observed in the HPV infections and squamous cells carcinoma (SCC). The Ck14, p53, p21 and Bcl-2 proteins were found modified in the leukoplakia, oral lichen planus and cancer. Cytological alterations and positive immunolabeling or over-expression of Ck14 cytokeratine in the upper epithelial stratus should be indicator of malignant transformations as doing subsequence exams.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Case-Control Studies; Cytodiagnosis; Female; Humans; Immunohistochemistry; Keratins; Leukoplakia, Oral; Lichen Planus, Oral; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Proteins; Papillomaviridae; Prognosis; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins p21(ras); Tumor Suppressor Protein p53

Cytokeratins (CKs) are the intermediate filament proteins of the epithelium cells, which have become important markers of normal and abnormal cell differentiation. The goal of the present study was to investigate the expression pattern of CK 10, 13, 14 and 16 in oral verrucous carcinoma (OVC) and oral squamous papilloma (OSP).. Formalin-fixed paraffin-embedded sections from eight cases of each lesion were assessed. Immunohistochemistry was carried out using streptoavidin-biotin complex method.. In OVC, CK 10 was expressed in suprabasal to superficial layers whereas in OSP mainly in superficial layer. CK 13 was detected in prickle and superficial cells in most cases of OVC and in suprabasal to superficial cells of OSP. All the cell layers of OVC reacted positively for CK 14 while basal and suprabasal layers of OSP were more pronounced for CK 14. Finally, CK 16 was observed in suprabasal to superficial layer in OVC and the majority cases in OSP showed only superficial reactive cells.. CK 10, 13, 14 and 16 immunohistochemical profile emphasis the biological behavior of the studied lesions and confirm the use of these proteins as markers of differentiation.

Topics: Carcinoma, Verrucous; Cell Proliferation; Cell Transformation, Neoplastic; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratinocytes; Keratins; Mouth Neoplasms; Papilloma

Cancer cells produce parathyroid hormone-related protein (PTHrP) in the early phase of malignancy development, before hypercalcemia occurs. The relationship between PTHrP and the clinicopathologic features of oral squamous cell carcinoma is poorly understood. We studied 60 patients (43 men, 17 women; mean age, 64.8 +/- 11.2 years) with primary oral squamous cell carcinoma, from whom pretreatment biopsy specimens were obtained. We examined the relationship among immunohistochemical PTHrP expression, serum PTHrP levels, clinical characteristics of the tumor, and histopathologic aspects of the tumor. The mean calcium concentration for the 60 patients was 9.1 +/- 0.4 mg/dl. No patients had laboratory evidence of hypercalcemia before treatment. Six patients had serum levels of C-terminal (C)-PTHrP higher than the normal level of 55.3 pmol/l. There were no significant differences in serum C-PTHrP levels according to TNM stages. Abundant positive immunoreactivity for anti-PTHrP (1-34) antibody was recognized diffusely in the whole cytoplasm of many tumor cells. Anti-PTHrP (38-64) antibody staining tended to localize as small granules in the cytoplasm, especially close to the nuclear periphery. There was no correlation between the serum C-PTHrP concentration and the intensity of either immunostain. The intensity of PTHrP was proportionally related to the degree of differentiation or extent of keratinization (P < 0.05) and the histologic malignancy grade of the tumor (P < 0.05), when using antibody against PTHrP (1-34), but not when using antibody against PTHrP (38-64). Serum C-PTHrP levels did not correlate with the intensity of cellular PTHrP expression and characteristics of the tumor at the initial patient visit. The fragment that includes PTHrP (1-34) may be involved in the differentiation of oral squamous cell carcinoma. The differences between immunoreactivities may have been due to differing tissue malignancies and the use of different antibodies. The results suggest the need for caution when interpreting immunoreactivities of PTHrP in malignancies.

Topics: Antibodies, Monoclonal; Biopsy; Calcium; Carcinoma, Squamous Cell; Cell Differentiation; Cell Nucleus; Cytoplasm; Cytoplasmic Granules; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Proteins; Neoplasm Staging; Parathyroid Hormone; Parathyroid Hormone-Related Protein; Peptide Fragments

We have previously reported that significantly higher levels of Keratin 14 (Ker-14) was observed in oral squamous cell carcinoma (OSCC) and severely dysplastic tissues, whereas this expression was reversed in hyperplasia and in mild to moderate dysplasia. In this study, the mechanism of Keratin 14 activation in oral squamous cell carcinoma (OSCC) cell lines (HSC-2, HSC-3 and Ca9-22) was investigated. Reporter analysis demonstrated that an upstream region (-1759/-1629) accounted for efficient promoter activity. Furthermore, electromobility sift and supershift assay demonstrated that interactions of the SP-1/SP-3 complex at the elements resided in -1737/-1702 and -1680/-1652 and may be essential for this activation in OSCC cells.

Topics: Binding Sites; Carcinoma, Squamous Cell; Cell Line, Tumor; Electrophoretic Mobility Shift Assay; Gene Expression Regulation, Neoplastic; Humans; Keratin-14; Keratins; Luciferases; Molecular Sequence Data; Mouth Neoplasms; Mutation; Oligonucleotide Probes; Promoter Regions, Genetic; Protein Binding; Recombinant Fusion Proteins; Regulatory Sequences, Nucleic Acid; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sp1 Transcription Factor; Sp3 Transcription Factor; Transcription, Genetic; Transfection

To validate lymphatic mapping combined with sentinel lymph node biopsy as a staging procedure, and to evaluate the possible clinical implications of added oblique lymphoscintigraphy and/or tomography and test the intra- and interobserver reproducibility of lymphoscintigraphy.. Forty patients (17 F and 23 M, aged 32-90) with 24 T1 and 16 T2 squamous cell carcinoma of the oral cavity. Planar lymphoscintigraphy, emission and transmission tomography were performed. Detection and excision of the sentinel nodes were guided by a gamma probe. The sentinel nodes were step-sectioning and stained with hematoxylin and eosin and cytokeratin (CK 1). Histology and follow-up were used as "gold standard". Tumor location, number of sentinel lymph nodes, metastasis, and recurrences were registered. Two observers evaluated the lymphoscintigraphic images to assess the inter-rater agreement.. Eleven (28%) patients were upstaged. The sentinel lymph node identification rate was 97.5%. Sentinel lymph node biopsy significantly differentiated between patients with or without lymph node metastasis (P = 0.001). Lymphatic mapping revealed 124 hotspots and 144 hot lymph nodes were removed by sentinel lymph node biopsy. Three patients developed a lymph node recurrence close to the primary tumor site during follow-up. Added oblique lymphoscintigraphic images and/or tomography revealed extra hotspots in 15/40 (38%) patients. In 4/40 (10%), extra contralateral hotspots were detected.. Sentinel lymph node biopsy upstaged 28% of the patients. Sentinel lymph nodes close to the primary tumor were difficult to find. Added oblique planar images and/or tomographic images revealed extra clinical relevant hotspots in 38% of patients. Reproducibility proved excellent.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Coloring Agents; Female; Follow-Up Studies; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neoplasm Recurrence, Local; Neoplasm Staging; Observer Variation; Radiology, Interventional; Radiopharmaceuticals; Reproducibility of Results; Rhenium; Sentinel Lymph Node Biopsy; Technetium Compounds; Tomography, Emission-Computed, Single-Photon

Metastasis to cervical lymph nodes (LN) is significantly associated with the outcome of patients with oral cancer. To provide a useful method for the detection of micrometastases, we analyzed 115 LNs from 10 patients with oral cancer using real-time quantitative polymerase chain reaction (PCR) based on the expression of squamous cell carcinoma antigen (SCCA) and cytokeratin 13 (CK13). The sensitivity and quantification of this method were assessed by means of limited dilution of cultured oral cancer cells and a model of cervical LN-metastasis established by inoculating green fluorescent protein (GFP)-expressing cells into the tongue of nude mice. In both investigations, a few cancer cells were detected by real-time quantitative PCR, but not by conventional reverse transcription-PCR (RT-PCR). SCCA mRNA was detected at high levels in metastatic LNs. In contrast, 26 of the 30 control cervical LNs did not express the gene at all, and the rest showed fairly low levels. Of 108 histologically metastasis-negative LNs, 19 (17.6%) expressed SCCA mRNA levels higher than the cut-off value (1.0: mean expression of control LNs + 2SD). CK13 mRNA is not a suitable marker for the real-time PCR since it was detected frequently even in the control LNs. These findings suggest that genetic diagnosis by real-time quantitative PCR based on SCCA mRNA expression may be clinically useful for detecting occult tumor cells in cervical LNs.

Topics: Animals; Antigens, Neoplasm; Cell Line, Tumor; Gene Expression Regulation, Neoplastic; Green Fluorescent Proteins; Humans; Keratins; Luminescent Proteins; Lymphatic Metastasis; Mice; Mice, Nude; Microscopy, Fluorescence; Mouth Neoplasms; Neoplasm Transplantation; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Serpins; Transfection; Transplantation, Heterologous

Squamous cell carcinoma of the oral cavity is one of the most common human neoplasms, and prevention of these carcinomas requires a better understanding of the carcinogenesis process and a model system in which cancer chemoprevention agents can be tested. We have developed a mouse model using the carcinogen 4-nitroquinoline 1-oxide (4-NQO) in the drinking water to induce tumorigenesis in the mouse oral cavity.. 4-NQO was delivered by tongue painting or drinking water to two mouse strains, CBA and C57Bl/6. The incidences of oral cavity carcinogenesis were then compared. In addition, we examined the expression of some of the molecular markers associated with the process of human oral cavity and esophageal carcinogenesis, such as keratin (K) 1, K14, p16, and epidermal growth factor receptor, by immunohistochemistry.. After treatment with 4-NQO in the drinking water, massive tumors were observed on the tongues of both CBA and C57Bl/6 female mice. Pathological analyses indicated that flat squamous dysplasias, exophytic papillary squamous tumors (papillomas), and invasive squamous cell carcinomas were present. Immunohistochemistry analyses showed that 4-NQO changed the expression patterns of the intermediate filament proteins K14 and K1. K14 was expressed in the epithelial suprabasal layers, in addition to the basal layer, in tongues from carcinogen-treated animals. In contrast, control animals expressed K14 only in the basal layer. Moreover, we observed more bromodeoxyuridine staining in the tongue epithelia of 4-NQO-treated mice. Reduced expression of the cell cycle inhibitor, p16, was observed, whereas 4-NQO treatment caused an increase in epidermal growth factor receptor expression in the mouse tongues. Interestingly, similar features of carcinogenesis, including multiple, large (up to 0.5 cm) exophytic papillary squamous tumors and invasive squamous cell carcinomas, increased bromodeoxyuridine staining, and increased K14 expression, were also observed in the esophagi of 4-NQO-treated mice. However, no tumors were observed in the remainder of digestive tract (including the forestomach, intestine, and colon) or in the lungs or livers of 4-NQO-treated mice. These results indicate that this murine 4-NQO-induced oral and esophageal carcinogenesis model simulates many aspects of human oral cavity and esophageal carcinogenesis.. The availability of this mouse model should permit analysis of oral cavity and esophageal cancer development in various mutant and transgenic mouse strains. This model will also allow testing of cancer chemopreventive drugs in various transgenic mouse strains.

Topics: 4-Nitroquinoline-1-oxide; Animals; Bromodeoxyuridine; Carcinogens; Carcinoma, Papillary; Carcinoma, Squamous Cell; Cyclin-Dependent Kinase Inhibitor p16; Disease Models, Animal; ErbB Receptors; Esophageal Neoplasms; Female; Immunoenzyme Techniques; Keratin-14; Keratins; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Mouth Neoplasms; Neoplasm Invasiveness; Tongue Neoplasms

We examined the effect of stable transfection of dominant negative TbetaR-II (dn TbetaR-II) cDNA in a human oral carcinoma cell line that contained normal Ras and was growth inhibited by TGF-beta1. Two clonal cell lines containing dn TbetaR-II were isolated and compared to the vector-only control and parent cell line. The treatment of cells with exogenous TGF-beta1 resulted in a decrease in ligand-induced growth inhibition and loss of c-myc downregulation in test cells compared to controls; transcriptional activation of certain genes including fra-1 and collagenase was retained. Cells containing dn TbetaR-II grew faster in monolayer culture, expressed less keratin 10 and exhibited increased motility and invasion in vitro compared to control cell lines. Endogenous TGF-beta1 production and the regulation of MMP-2 and MMP-9 by TGF-beta1 remained unchanged. After orthotopic transplantation to the floor of the mouth in athymic mice, cells containing dn TbetaR-II formed comparable numbers of primary tumours at the site of inoculation as controls but the tumours were less differentiated as demonstrated by the absence of keratin 10 immunostaining. Further, metastatic dissemination to the lungs and lymphatics was more evident in grafts of cells containing dn TbetaR-II than controls. Taken together, the results demonstrate that attenuation of TGF-beta signalling through transfection of dn TbetaR-II cDNA leads to an enhanced growth rate, a loss of tumour cell differentiation and an increase in migration and invasion, characteristics that corresponded to the development of the metastatic phenotype.

Topics: Cell Differentiation; Cell Division; Cell Line, Tumor; Cell Movement; Humans; Keratin-10; Keratinocytes; Keratins; Lung Neoplasms; Mouth Neoplasms; Neoplasm Metastasis; Phenotype; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Signal Transduction; Transforming Growth Factor alpha

Population-based oral cancer screening appears to be a promising health promotion strategy (especially in high-risk individuals) with significant increases in quality-adjusted life years saved. However, the current protocol, conventional visual inspection, and palpation of oral soft tissues for the early detection of pre-malignant or malignant changes, appears to be deficient. The adjunctive application of technology to highlight such lesions may increase the diagnostic yield. The purpose of this pilot study was to collect data, which might support the hypothesis that oral soft tissues exhibit features similar to the cervical epithelium following an acetic acid wash and visual inspection under chemiluminescent illumination. The data provides strong evidence to support the hypothesis. Epithelium with hyperkeratinization, hyperparakeratinization, and/or chronic inflammatory infiltrate reflects the diffuse, low-level, blue-white chemiluminescent light more strongly and appears amplified. Similarly, epithelium with an altered nuclear-cytoplasmic ratio also reflects the diffuse, low-level, blue-white chemiluminescent light. In such cases, the lesions become clinically discernible and appear "acetowhite." Large-scale studies are required to further refine issues related to the selectivity and specificity of the technology.

Topics: Acetic Acid; Adolescent; Adult; Aged; Cell Nucleus; Cytoplasm; Epithelium; Female; Humans; Hyperplasia; Indicators and Reagents; Keratins; Leukoplakia, Oral; Luminescent Measurements; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Pilot Projects; Precancerous Conditions; Sensitivity and Specificity

This study examines differences between cultures of normal human oral epithelial cells and two squamous cell carcinoma cell lines (SCC15 and SCC25) in the expression of structural proteins, adhesion molecules, plasma membrane lipid composition, and intercellular junctions. Based on immunocytochemistry, most normal cell cultures appeared to express more E-cadherin, integrin beta-1, cytokeratin (CK) 14, CK19, and involucrin than SCC cultures. By Western blot analysis, normal cultures expressing high levels of E-cadherin also expressed high levels of involucrin and low levels of CK19. Both SCC cultures demonstrated lower expression of E-cadherin and involucrin, whereas only SCC15 cells showed high levels of CK19. Expression of beta-catenin, an E-cadherin associated protein with potential oncogene function, did not vary among normal and SCC cells. Proportions of saturated fatty acids quantified by thin layer chromatography were higher in the normal cell cultures, than in both SCC cell lines. No morphological differences were evident by transmission electron microscopy (TEM) between normal and SCC cell-cell intercellular junctions. Although no quantitation was attempted, observation suggested that normal cells form more intercellular junctions (TEM observation) and larger intercellular bridges (SEM observation) compared to both SCC cell lines. Of the factors examined, main variations between cultures of normal oral epithelium and the two SCC cell lines examined include the expression of structural and adhesion proteins, lipid composition, and intercellular junctions. The extent of the differences varies according to the stage of terminal differentiation demonstrated by the normal cell cultures.

Topics: Biomarkers, Tumor; Blotting, Western; Cadherins; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line, Tumor; Cells, Cultured; Fatty Acids; Gingiva; Humans; Immunohistochemistry; Integrin beta1; Keratinocytes; Keratins; Microscopy, Electron; Mouth Neoplasms; Protein Precursors

The cytokeratin (CK) pair 8 and 18 is normally expressed in all simple epithelia. This pair is not normally seen in stratified epithelial cells. Squamous cell carcinomas derived from stratified epithelia show anomalous expression of this CK pair. It is not known whether CKs 8 and 18 in any way contribute to the malignant phenotype of these cells. We used an immortalised, nontransformed human foetal buccal mucosa (FBM) cell line that expresses significantly higher amounts of CK18 compared to CK8. FBM cells were transfected with the full-length CK8 gene to study the role of CKs 8 and 18 in malignant transformation. Clones with higher expression of CK8 compared to untransfected FBM cells were studied for changes in their phenotypic characteristics. Immunofluorescence studies using antibodies to CKs 8 and 18 revealed well-decorated filaments throughout the cytoplasm in CK8 gene-transfected cells vs. untransfected FBM cells. Transmission images showed that FBM cells were isolated while transfected cells were in groups of well-spread cells with cellular projections. Transfected cells were independent of growth supplement requirements and showed anchorage-independent growth in soft agar assay and significantly reduced doubling time compared to nontransfected FBM cells. DNA flow-cytometric studies revealed increased DNA content and prolonged S phase in transfected clones vs. FBM cells. Injection of cells s.c. obtained from soft agar colonies developed from 2 of the clones resulted in tumour formation at the site of injection. In both cases, lung metastasis was also seen. Thus, in conclusion, it appears that increased expression of CK8 in some way changes the phenotypic characteristics of stratified epithelial cells, resulting in malignant transformation.

Topics: Animals; Carcinoma, Squamous Cell; Cell Cycle; Cell Division; Cell Line; Cell Transformation, Neoplastic; DNA; Epithelial Cells; Flow Cytometry; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Humans; Keratins; Mice; Mice, Nude; Mouth Mucosa; Mouth Neoplasms; Neoplasms, Experimental; Phenotype; Transfection; Transplantation, Heterologous

Mouse models for cancer represent powerful tools to analyze the causal role of genetic alterations in cancer development. We have developed a novel mouse model that allows the focal activation of mutations in stratified epithelia. Using this system, we demonstrate that activation of an oncogenic K-rasG12D allele in the oral cavity of the mouse induces oral tumor formation. The lesions that develop in these mice are classified as benign squamous papillomas. Interestingly, these tumors exhibit changes in the expression pattern of keratins similar to those observed in human premalignant oral tumors, which are reflective of early stages of tumorigenesis. These results demonstrate a causal role for oncogenic K-ras in oral tumor development. The inducible nature of this model also makes it an ideal system to study cooperative interactions between mutations in oncogenes and/or tumor suppressor genes that are similar to those observed in human tumors. To our knowledge, this is the first reported inducible mouse model for oral cancer.

Topics: Animals; Blotting, Western; Carcinoma, Squamous Cell; Epithelial Cells; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Genes, ras; Keratins; Mice; Mice, Knockout; Mouth Neoplasms; Mutation; Oncogene Protein p21(ras); Papilloma; Polymorphism, Restriction Fragment Length; Precancerous Conditions; Reverse Transcriptase Polymerase Chain Reaction

DNA microarrays are powerful tools for exploring gene expression and predicting disease state. However, since the number of variables (genes) typically exceeds the number of samples (tissue specimens), many potentially spurious genes may be selected for a predictor function. Principle component analysis (PCA) can greatly reduce the high-dimensional microarray data space while retaining most of the inherent variability. We propose a methodology that uses PCA to identify a predictor vector between two mutually exclusive and collectively exhaustive classes. By projecting the training set upon this vector a distribution of projections can be computed for each class. A log-likelihood ratio is then calculated for class membership. We used this methodology to classify 48 biopsy specimens as either oral squamous cell carcinoma or normal oral mucosa using oligonucleotide microarrays. The system was trained using a set of half the samples, and correctly predicted the membership of the other half. The three most highly positively and three most highly negative predictive genes were all keratins that are known markers of squamous cell carcinoma.

Topics: Biopsy; Carcinoma, Squamous Cell; Gene Expression Profiling; Genetic Markers; Humans; Keratins; Linear Models; Mouth Mucosa; Mouth Neoplasms; Oligonucleotide Array Sequence Analysis; Principal Component Analysis; RNA, Messenger

Squamous cell carcinoma (SCC) is the most common form of oral malignancy and is often preceded by premalignant lesions, some of which are more likely to progress to carcinoma than others. In this study, a panel of monoclonal antibodies (AE1/AE3, cytokeratin [CK] 14, Ki-67 and p53) is applied to 10 cases of human oral tissue in each of six categories to establish staining patterns indicative of which lesions are more likely to progress to malignancy. The six tissue categories are normal tissue; abnormal benign lesions; mild, moderate and severe dysplasia; and SCC. A statistical analysis of Ki-67 and p53 immunoexpression is performed. The results showed that AE1/AE3 and CK 14 expression was reduced as a late event in oral carcinogenesis, particularly in poorly differentiated SCC. Expression of Ki-67 and p53 proved to be a weak but statistically significant predictor of malignant progression in oral tissue.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Mouth Neoplasms; Neoplasm Proteins; Precancerous Conditions; Tumor Suppressor Protein p53

To investigate the relationship of vascular endothelial growth factor C (VEGF-C) and induced nitride oxide synthesizase (iNOS) expression in lymph node micrometastasis of oral squamous cell carcinoma.. Samples were obtained from 47 cases of oral squamous cell carcinoma and 15 cases with normal oral mucosa, VEGF-C and iNOS mRNA expression were detected by RT-PCR method. Lymph node micrometastasis of 10 normal lymph nodes and 355 lymph nodes from 47 cases of oral squamous cell carcinoma was detected with immunohistochemical reaction in cytokeratin antibody.. The percentages in tumors with higher expression were 57.4% for VEGF-C, 68.1% for iNOS (P < 0.05). They were significantly higher than that of normal groups. Significant positive relationship was found between VEGF-C and iNOS (P < 0.01). The positive rate of cytokeratin (CK) was 48.9%. Significant positive relationship was found between VEGF-C and CK, iNOS and CK (P < 0.01). The expression rates of CK in positive group of VEGF-C and iNOS were 63.0%, 65.6% respectively, and were significant higher than negative groups.. Expression of VEGF-C and iNOS in lymph node micrometastasis of oral squamous cell carcinoma is significant related.

Topics: Carcinoma, Squamous Cell; Humans; Keratins; Lymphatic Metastasis; Mouth Neoplasms; Neoplasm Micrometastasis; Nitric Oxide Synthase Type II; Vascular Endothelial Growth Factor C

The value of histological grading was examined with emphasis on reliability of assessment in 102 cases of intraoral squamous cell carcinoma from Northern Ireland with known outcome.. Two pathologists independently graded the invasive tumour front blinded to the stage and outcome.. Intraobserver agreement was acceptable but interobserver agreement was not satisfactory. The degree of keratinisation was assessed most consistently while nuclear polymorphism was the least reliable feature. Multivariate survival analysis showed that the total grading score was associated with overall survival while the pattern of tumour invasion was the most valuable feature in estimating regional lymph node involvement. The number of positive lymph nodes was strongly associated with regional relapse, while the treatment modality and status of the surgical margins correlated with local relapse.. Grading of selected features in OSCC is reliable and can facilitate treatment planning.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lymphatic Metastasis; Male; Middle Aged; Mitotic Index; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Staging; Observer Variation; Polymorphism, Genetic; Prognosis; Proportional Hazards Models; Reproducibility of Results; Survival Analysis

The frequency of amyloid deposits associated with squamous cell carcinoma (SCC) and dysplasia in the oral cavity, pharynx and larynx was examined. In addition, the origin of amyloid proteins by immunohistochemical staining with a panel of anticytokeratin monoclonal antibodies was investigated. Amyloid deposits were found in eight of 73 (11.0%) SCC and one of seven (14.3%) dysplasias in the oral cavity, in eight of 22 (36.4%) SCC and zero of two (0%) dysplasias in the pharynx, and in 22 of 37 (59.5%) SCC and four of 10 (40.0%) dysplasias in the larynx. Eight of 12 different cytokeratin (CK) antibodies reacted with these deposits: 34 beta E12 (CK1, -5, -10, -14) reacted with amyloid deposits in 19 of 19 cases (100%), LL002 (CK14) in eight of 18 cases (44.4%), MNF116 (CK5, -6, -8, -17) in eight of 19 cases (42.1%), D5/16B4 (CK5, -6) in five of 18 cases (27.8%), DE-K10 (CK10) in four of 17 cases (23.5%), RCK108 (CK19) in three of 18 cases (16.7%), 34 beta B4 (CK1) in three of 19 cases (15.8%) and AE8 (CK13) in two of 17 cases (11.8%). These antibodies always reacted with the cytoplasm of squamous cell lesions. Amyloid deposits in two cases contained a CK5 and CK14 pair, and in another two cases they contained both a CK5 and CK14 pair, and a CK1 and CK10 pair. Anti-CK antibodies, including OV-TL12/30 (CK7), c-51 (CK8), DC10 (CK18) and IT-Ks20.8 (CK20) did not react with the amyloid deposits. We conclude that the amyloid deposits associated with SCC or dysplasia in the oral cavity, pharynx or larynx were derived from CK of cancer cells and that some amyloid deposits might be assembled by two or more different CK.

Topics: Adult; Aged; Aged, 80 and over; Amyloid; Amyloidosis; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Pharyngeal Neoplasms; Precancerous Conditions

White sponge naevus (WSN) is a rare, autosomal dominant disorder that predominantly affects noncornified stratified squamous epithelia, most commonly the buccal mucosa. Clinically, WSN manifests as thickened spongy mucosa with a white opalescent tint in the mouth and may be confused with other disorders that cause white lesions on oral mucosa. Recent studies have identified pathogenic mutations in KRT4 and KRT13, the genes encoding mucosa-specific keratins, in WSN.. To search for possible mutations in KRT4 and KRT13.. We report a case of WSN in a young man who presented with diffuse irregular whitish plaques involving the buccal and gingival mucosae and the tongue. Results Pathologically, the affected mucosa showed epithelial thickening, parakeratosis and extensive vacuolization of the suprabasal keratinocytes. Mutation analysis revealed a heterozygous missense mutation 1345G-->A in KRT4, predicting an amino acid change, E449K, in the 2B domain of the K4 polypeptide.. We report the first mutation analysis of a Taiwanese patient with WSN. Potentially this novel mutation could disrupt the stability of keratin filaments and result in WSN.

Topics: Adult; Amino Acid Sequence; Base Sequence; Humans; Keratins; Male; Molecular Sequence Data; Mouth Mucosa; Mouth Neoplasms; Mutation; Nevus; Pedigree; Polymerase Chain Reaction; Skin Neoplasms

Cytokeratins (CK) are the epithelia specific intermediate filament proteins. We have shown consistent non-expression of CK-5 protein in human oral pre-cancer and cancer, in earlier studies. To investigate whether non-expression of CK-5 protein is the result of transcriptional or translational block and to evaluate the possibility if CK-5 non-expression can be used as a marker for early diagnosis of tobacco related oral cancer, RT-PCR using CK-5 specific primers was conducted. Out of 36 precancerous lesions and 29 squamous cell carcinomas (SCC) of buccal mucosa (BM) samples studied, 11 and 13 samples respectively of precancer and SCC did not show CK-5 product in RT-PCR. Down regulation of CK-5 mRNA expression was also observed in some samples. Thus, in conclusion, our results have shown that CK-5 non-expression is the result of transcriptional block. We proposed CK-5 non-expression as a potential marker for the early diagnosis of tobacco related oral cancer.

Topics: Carcinoma, Squamous Cell; Case-Control Studies; Cheek; Gene Expression Regulation; Genetic Markers; Humans; Keratins; Leukoplakia, Oral; Mouth Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; Tobacco Use Disorder; Tobacco, Smokeless; Transcription, Genetic

Basaloid squamous carcinoma (BSC) is an aggressive variant of squamous cell carcinoma (SCC) with a predilection for the upper aerodigestive tract. In the English literature, approximately 40 cases of BSC have been described in the oral cavity. BSC has frequently been confused with adenoid cystic carcinoma (ACC), basal cell adenocarcinoma, and undifferentiated SCC. The purpose of the investigation was to examine the histological features and immunohistochemical expression of differentiation-related substances, including cytokeratin (CK) subtypes, vimentin, S-100, chromogranin, laminin, and type IV collagen, for the characterization of biological features of these tumours. We studied three cases of BSC of the oral cavity, three cases of ACC, and one case of basal cell adenocarcinoma. Well-differentiated and undifferentiated SCCs were also studied for comparison. The BSCs showed many histopathologic similarities to cases previously reported. Among the CK subtypes analyzed, CK14 was the only subtype expressed by all basaloid cells of BSC. Potentially useful for the differential diagnosis was the finding of CKs 7 and 19 expression in the basaloid cells of ACC, and CKs 7 and 8 in basal cell adenocarcinoma. In BSCs, laminin and type IV collagen were found in the microcystic spaces between basaloid cells, but neither ACCs nor basal cell adenocarcinoma showed this feature. These data suggest that immunohistochemical findings are helpful in distinguishing BSC of the oral cavity from other histopathologically similar tumours.

Topics: Adenocarcinoma; Carcinoma, Adenoid Cystic; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Mouth Neoplasms

We evaluated the importance of several tumor factors related to predicting the presence of occult metastases in the oral cavity.. Retrospective case study.. The study comprises 29 patients treated at the Department of Otorhinolaryngology (University of Trieste, Cattinara Hospital, Trieste, Italy) between January 1990 and December 2000, who had T1-T2 carcinoma of the oral cavity that had or had not extended to the oropharynx and were clinically evaluated as N0 neck. The patients all underwent surgery with removal of tumor and neck dissection. Four tumor-related parameters were examined with the aim of evaluating their predictivity of metastasis: tumor class, degree of keratinization, degree of differentiation according to Brooler's histopathological grading, and invasive cell grading (ICG). With the exception of tumor class, these parameters were evaluated both in the biopsy and in the surgical specimen and the findings were then compared. We evaluated existing correlations between each individual parameter and the histopathological presence of micrometastases (pN+) and extracapsular spread revealed when specimens from the neck were examined.. There was a highly significant correlation between ICG equal to or greater than 13 (range, 5-20) and the presence of occult metastases (P =.0017). On the basis of our findings, the ICG parameter correctly identified 9 of 10 (pN+) patients and could have reduced overtreatment from 65.5% to 17.2% in histopathologically negative necks (pN0).. It would appear that with a delay in programming a neck dissection so as to consider ICG in combination with thickness, as in seven recent patients, identification of locoregional occult metastases (pN+) might be more precise.

Topics: Adult; Aged; Female; Humans; Keratins; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neck Dissection; Neoplasm Invasiveness; Neoplasm Staging; Oropharyngeal Neoplasms; Prognosis; Retrospective Studies

In the literature, few studies based on clinical and histological evaluation analyze skin structural changes after transplantation to the oral cavity. Ten patients affected by squamous cell carcinoma of the oral cavity who were reconstructed with a free forearm flap were included in the current study to analyze skin alterations. The authors performed a histological and ultrastructural evaluation of skin samples from the free forearm flap before transplantation and 18 months after intraoral reconstruction. They analyzed cytokeratin and involucrin distribution, which represent markers of maturation and differentiation of epithelia. The aim of this study was to demonstrate whether skin "mucosalization" occurs. They found that the skin undergoes some morphological changes induced by the intraoral environment. Cytokeratin and involucrin distribution is mostly unchanged. This aspect is in favor of skin structure preservation. Thus, they found that "mucosalization" of the skin is not evident after 18 months.

Topics: Aged; Carcinoma, Squamous Cell; Female; Humans; Immunohistochemistry; Keratins; Male; Microscopy, Electron, Scanning; Middle Aged; Mouth; Mouth Neoplasms; Protein Precursors; Skin; Skin Transplantation; Surgical Flaps

To investigate the changes of cytokeratin 19 during oral carcinogenesis.. 53 specimens including normal oral mucosa, oral epithelial hyperplasia, oral epithelial dysplasia and oral squamous cell carcinoma were investigated by immunohistochemistry, SDS-PAGE and Western blotting.. CK19 was detectable in suprabasal cell layers in epithelial dysplasia and in oral cancer, especially in poor-differentiated cancerous cells. With the lesions getting worse, the positive rate, the intensity and the constituent ratio of CK19 raised significantly.. The results suggest that the CK19 expression in suprabasal cell layers of oral mucosa can be used as a marker of diagnosis of oral precancerous lesions and CK19 expression is the initial events during oral carcinogenesis.

Topics: Adult; Aged; Blotting, Western; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions

A tumour-secreted cytokine autocrine motility factor (AMF) induces in vivo invasion and metastasis, and in vitro tumour cell motility by a signal transduction through interaction with its cell surface receptor gp78. In this report, we investigated the characterization of a high-metastatic human oral squamous cell carcinoma (SCC) cell line LMF4 and low-metastatic HSC-3 in comparison with non-metastatic HSC-2 and HSC-4. Morphological and motility analyses revealed LMF4 cells to have the highest motile activity among those cells. However, LMF4 cells shared the similar features with HSC-3: high level secretion of AMF, enhancement of gp78 expression, co-expression of vimentin and cytokeratin, although LMF4 cells showed twice as high motile reactivity as HSC-3. The only difference was that LMF4 had twice as high amount of low-affinity receptor(s) as HSC-3, shown by Scatchard analysis.

Topics: Blotting, Northern; Blotting, Western; Carcinoma, Squamous Cell; Cell Movement; Confidence Intervals; Electrophoresis, Polyacrylamide Gel; Glucose-6-Phosphate Isomerase; Humans; Keratins; Mouth Neoplasms; Neoplasm Invasiveness; Receptors, Autocrine Motility Factor; Receptors, Cytokine; Tumor Cells, Cultured; Ubiquitin-Protein Ligases; Vimentin

The ability of oral squamous cell carcinoma to metastasize to lymph nodes does not always show a relationship with clinical staging. The aim of this study was to attempt to define a trend for predictive histopathologic and/or molecular biomarkers in the development of nodal metastasis by analyzing 2 clinically extreme groups.. Patients with small primary tumors (T1, T2) with lymph node metastasis and patients with large primary tumors (T3, T4) without metastatic disease were identified among 315 consecutive cases of primary oral squamous cell carcinoma. Group comparisons were made with use of a Mann-Whitney test, and associations among the variables were assessed with nonparametric and parametric correlational analyses.. The degree of keratinization was significantly less in primary tumors with lymph node metastasis (P < or = .01). The degree of keratinization was significantly associated with nuclear pleomorphism (P =.02), number of mitoses (P =.02), stage of invasion (P =.002), and p53 expression (P =.04), independent of clinical stage of the tumor. Other microscopic features and immunohistochemical markers did not differ significantly between the groups (P >.05).. These data indicate that there still is no single predictive parameter superior to the degree of keratinization to identify patients at risk for the development of regional metastasis.

Topics: Adult; Aged; Biomarkers; Carcinoma, Squamous Cell; Cell Nucleus; Female; Forecasting; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratins; Lymphatic Metastasis; Male; Middle Aged; Mitosis; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Staging; Proliferating Cell Nuclear Antigen; Proto-Oncogene Proteins c-bcl-2; Receptor, ErbB-2; Risk Factors; Statistics as Topic; Statistics, Nonparametric; Tumor Suppressor Protein p53

Cytokeratins (also known as keratins (K)) are members of the family of intermediate filaments and form major components of the mammalian epithelial cell cytoskeleton. Cytokeratins have emerged as reliable cellular markers of oral cancer development and chemoprevention because of their abundance, stability and high antigenicity.. We investigated the effect of aqueous garlic extract on cytokeratin expression during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. Hamsters were divided into four groups of six animals. Animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin, on the right buccal pouches, three times a week for 14 weeks. Group 2 animals were painted with DMBA as in group 1 and also received 250 mg/kg body weight aqueous garlic extract orally on alternate days to the DMBA application. Group 3 animals received garlic extract only, as in group 2. Group 4 animals received neither DMBA nor garlic extract and served as the control. The hamsters were killed after an experimental period of 14 weeks.. Cytokeratin expression was studied using human monoclonal antibodies AE1 and AE3, which react with type I and II keratins. In DMBA-induced squamous cell carcinomas, decreased expression of high molecular weight keratins was observed. Administration of garlic extract to animals painted with DMBA suppressed HBP carcinomas and restored normal cytokeratin expression.. The results of the present study suggest that inhibition of HBP carcinogenesis by garlic may be due to its regulatory effects on differentiation, tumour invasiveness, migratory and metastatic potential. We suggest that one of the mechanisms of tumour inhibition by garlic is an influence on cellular differentiation.

Topics: Animals; Antibodies, Monoclonal; Antineoplastic Agents, Phytogenic; Blotting, Western; Carcinoma, Squamous Cell; Cell Differentiation; Cheek; Chemoprevention; Cricetinae; Disulfides; Electrophoresis, Polyacrylamide Gel; Garlic; Humans; Keratins; Male; Mesocricetus; Mouth Neoplasms; Phytotherapy; Random Allocation; Sulfinic Acids

A 70-year-old man was admitted to hospital because of oral and cervical masses. Computed tomographic scanning revealed a lobulated mass lesion in the retropharyngeal region, with a protruding extension in the oral cavity and with destruction of the second cervical vertebra. A biopsy was performed under the diagnosis of a retropharyngeal tumor. Histologically, this lesion was composed of vacuolated tumor cells in a solid or cord-like arrangement, with an abundant myxoid matrix. Immunohistochemically, the tumor cells were positive for pancytokeratin, epithelial membrane antigen and S-100 protein. The tumor was diagnosed as chordoma. Chordoma presenting as an intra-oral mass lesion is very rare.

Topics: Aged; Cervical Vertebrae; Chordoma; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Male; Mouth Neoplasms; Mucin-1; S100 Proteins; Spinal Neoplasms

Our study examined the expression of AP-1 family members in keratinocytes derived from the rat-4NQO model of oral carcinogenesis in which extremes of epithelial differentiation and tumour cell aggressiveness are evident. The constitutive expression of JunB was diminished in the undifferentiated, more aggressive tumour phenotype compared with the well-differentiated, less aggressive keratinocytes, whereas the expression of other AP-1 family members (c-jun, junD, c-fos, fra1, fra2 and fosB) was either very weak or variable. After transfection of the undifferentiated keratinocytes with junB cDNA, clonal populations were isolated that expressed similar levels of JunB protein as the well-differentiated cells. Both untransfected and transfected cell lines were keratin negative and vimentin positive. Increased expression of JunB in the transfected cells resulted in up-regulation of c-Jun and Fra1 and an enhanced AP-1 activity as demonstrated by transcriptional activation of the prototypic AP-1 dependent promoter, MMP-1. JunB transfected cells grew more quickly than vector-only controls and were refractory to the growth inhibitory effects of TGF-beta1. Over-expression of JunB resulted in the elevated expression of the AP-1 dependent proteinase, MMP-9, whereas the expression of the AP-1 independent enzyme, MMP-2, was unaffected. JunB transfected keratinocytes were highly invasive in an in vitro assay of tumour cell invasion compared with vector controls. The results indicate that increased expression of JunB above baseline levels in undifferentiated rat keratinocytes does not alter epithelial differentiation but enhances the malignant phenotype in vitro, possibly by altering the dynamics of the AP-1 complex.

Topics: 3T3 Cells; 4-Nitroquinoline-1-oxide; Animals; Bacterial Proteins; Blotting, Northern; Blotting, Western; Carcinogens; Cell Differentiation; Cell Division; Cell Movement; Cells, Cultured; DNA-Binding Proteins; DNA, Complementary; Fos-Related Antigen-2; Genes, Reporter; Genetic Vectors; Keratinocytes; Keratins; Luciferases; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Mice; Mouth Neoplasms; Neoplasms, Experimental; Phenotype; Plasmids; Promoter Regions, Genetic; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-jun; Rats; Time Factors; Transcription Factor AP-1; Transcription Factors; Transcriptional Activation; Transfection; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Cells, Cultured; Up-Regulation; Vimentin

The aim of the study was to investigate the differentiation-specific keratins (K4, K13, K1 and K10) in oral epithelial dysplasia and squamous cell carcinoma (SCC). Alterations in keratin gene expression were determined by in situ hybridization using 35S-labeled riboprobes and immunohistochemistry with monoclonal antibodies. In mild dysplasia, both sets of differentiation keratins were expressed in the same group of cells but in moderate lesions, expression of K4 and K13 was reduced in the presence of enhanced K1 and K10 synthesis. In severe dysplasia, neither mRNAs nor proteins were detected. In tumor islands of well and moderately differentiated SCCs, the K4/K13 complex was co-expressed with K1/K10, but in poorly differentiated carcinomas, differentiation keratins were absent. Consequently, mild oral epithelial dysplasia and well differentiated SCC retain an essentially normal pattern of keratin gene expression and hence epithelial differentiation while in severe dysplasia and poorly differentiated SCC keratin gene expression reflects the gross changes in epithelial differentiation and maturation.

Topics: Carcinoma, Squamous Cell; Cell Differentiation; Cheek; Epithelium; Gene Expression; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; RNA, Messenger; Staining and Labeling

Human beta-defensin(hBD)-2, an antimicrobial peptide, is produced by various epithelial cells. Because hBD-2 expression in the oral epithelium has not been assessed, we investigated its localization in normal oral epithelium and epithelial lesions. hBD-2 expression was studied using immunohistochemistry and in situ hybridization on formalin-fixed, paraffin-embedded tissue sections from 30 cases of squamous cell carcinoma and 6 cases of leukoplakia. Immunostaining for hBD-2 was more intense in hyperkeratinized than in ortho- or non-keratinized epithelium. In contrast, signals for hBD-2 mRNA were frequently stronger in non-keratinized epithelium than in hyper- or ortho-keratinized epithelium. The results suggest that keratinization in oral epithelium plays an important role in the biological function of hBD-2 both at the mRNA level and in the retention of the peptide in the epithelium.

Topics: beta-Defensins; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Mouth Mucosa; Mouth Neoplasms; RNA, Messenger

Keratins have been extensively studied in tissues and cultured keratinocytes but limited information is available on epithelia reconstructed in vitro. The aim of this study was to examine keratin expression in organotypic epithelia with normal (NOK), immortalized (SVpgC2a) and malignant (SqCC/Y1) human buccal cells. Organotypic epithelia were derived from 10 days of culture at the air-liquid interface of collagen gels containing human oral fibroblasts using a standardized serum-free medium. Sections were stained immunohistochemically with selected mono-specific antibodies to a range of keratins. Organotypic epithelia showed sharp differences in keratin expression and distribution. K4/K13, K1/K10, K6/K16 were variably expressed in NOK and SqCC/Y1 but were not detected in SVpgC2a. K5 was expressed in all organotypic epithelia but K14 was absent in SVpgC2a. K7 and K8 showed variable expression while K18 was expressed uniformly in all epithelia. K19 was expressed consistently in NOK and K20 was distributed heterogeneously in SVpgC2a. Overall, organotypic cultures of normal keratinocytes express many of the same keratins as buccal mucosa. Further, the loss of keratins in SVpgC2a and their retention in SqCC/Y1 have several features in common with the respective keratin profile of oral epithelial dysplasia and well-differentiated oral squamous cell carcinoma. Although qualitative and quantitative differences exist compared to keratin expression in vivo, these cell lines in organotypic culture may serve in studies of the multi-step progression of oral cancer.

Topics: Carcinoma, Squamous Cell; Case-Control Studies; Epithelial Cells; Fibroblasts; Humans; Keratinocytes; Keratins; Mouth Mucosa; Mouth Neoplasms; Tumor Cells, Cultured; Up-Regulation

In an effort to come to a better understanding of human oral mucosal carcinogenesis, an animal model was used in which the carcinogen 4-nitroquinoline-1-oxide was applied to rat palatal mucosa for varying periods of time. Histological and histometric analyses showed that there were quantifiable differences in the palatal epithelium to which carcinogen had been applied in comparison with control tissue. Tissue recombination experiments, using various combinations of the palatal mucosa and analysed after recovery from transplantation to hypothymic BALB/c mice, showed that control epithelium recombined with connective tissue from carcinogen-treated mucosa was altered, indicating that the underlying connective tissue modified histomorphological aspects of the epithelium in the later stages of carcinogenesis.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinogens; Connective Tissue; Disease Models, Animal; Epithelium; Humans; Image Processing, Computer-Assisted; Keratins; Male; Mesoderm; Mice; Mice, Inbred BALB C; Mice, Nude; Mouth Mucosa; Mouth Neoplasms; Rats; Rats, Sprague-Dawley; Statistics, Nonparametric; Transplantation, Heterologous

To examine whether cancer cell dissemination results from incisional biopsy, we tried to detect squamous cell carcinoma (SCC) cells in peripheral blood before and after incisional biopsy by means of cytokeratin 19 (CK19) reverse-transcriptase polymerase chain reaction (RT-PCR). The study population consisted of 20 patients with oral SCC; 10 were given incisional biopsies followed by radical excision (the incisional biopsy group), and the remaining 10 were treated by excisional biopsy alone (the excisional biopsy group). Ten non-oral cancer patients with benign oral lesions served as controls. Five-ml blood aspirates collected before and after incision were used for CK19 RT-PCR. Two (20.0%) of 10 patients from the incisional biopsy group were positive for CK19 transcripts in their peripheral blood drained 15 min after incision. In contrast, CK19 transcript was not detected either in the excisional biopsy group or in controls. Surgical invasiveness for oral cancer, including incisional biopsy, causes dissemination of cancer cells into circulation, resulting in increased risk of metastasis.

Topics: Aged; Aged, 80 and over; Biopsy; Carcinoma, Squamous Cell; Case-Control Studies; DNA, Neoplasm; Female; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Seeding; Reverse Transcriptase Polymerase Chain Reaction

Cytokeratin (CK) 20 is a low molecular-weight intermediate filament reportedly expressed only by benign and malignant gastrointestinal epithelium, urothelium and Merkel cells. The main aims here were to map its expression in normal oral mucosa of humans and other mammals, and to determine whether it was expressed by abnormal human oral epithelium. Salivary and odontogenic epithelium were also analysed. An immunoperoxidase method was used on wax-embedded and cryostat sections. In addition, double-labelling experiments were undertaken to determine the association between CK 20 expression and that of CK 8/18 or S100 protein. Normal human oral mucosa from four sites, together with abdominal skin, was studied in autopsy samples from 32 individuals. CK 20-positive, basally situated, round or angular cells, consistent with Merkel cells, were recorded in 24/32 (75.0%) samples of mandibular gingiva, 25/32 (78.1%) samples of hard palate, 7/32 (21.9%) samples of buccal mucosa, 0/32 samples of lateral border of tongue, and 2/32 (6.3%) samples of abdominal skin. Double-labelling showed that all CK 20-positive Merkel cells also expressed CK 8/18 and S100. The only other cells to express CK 20 were human taste buds. There was no expression by dysplastic or invasive oral epithelium from biopsy samples. Colonic mucosa showed luminal-cell positivity in man, marmoset, ferret, rabbit and guinea-pig, but oral mucosa was universally negative in non-human species. It is concluded that in oral mucosa CK 20 is a specific marker of Merkel cells and taste buds, that Merkel cells are more frequently present in keratinized than non-keratinized oral mucosa, that CK 20-positive Merkel cells are also S100-positive, that there may be interspecies variations in CK 20 polypeptide composition and that, by contrast to urothelium, CK 20 has no value in the diagnosis of oral epithelial dysplasia.

Topics: Adult; Aged; Aged, 80 and over; Animals; Biomarkers, Tumor; Callithrix; Cats; Cattle; Cricetinae; Epithelium; Female; Ferrets; Gingiva; Guinea Pigs; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratins; Macaca mulatta; Male; Merkel Cells; Mesocricetus; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Odontogenic Cysts; Palate, Hard; Rabbits; Rats; S100 Proteins; Salivary Glands; Sheep; Skin; Swine; Taste Buds; Tongue

Diploid tumour cells regularly continue to progress after the development of aneuploid cell populations in head and neck squamous cell carcinomas. The coexistence of aneuploid clones with their diploid progenitor cells provides a unique opportunity to study the order of appearance of p53 mutation and aneuploidy in the same tumour. Multiparameter flow cytometry was therefore applied to 22 oral squamous cell carcinomas to simultaneously assess cellular DNA content and p53 protein expression on a single-cell basis. Concurrent measurements of cytokeratin expression served to identify tumour cells of epithelial origin. One of 5 diploid and 2 of 17 aneuploid carcinomas were p53-negative. For 15 p53-positive aneuploid tumours, overexpression of p53 protein was identified for the aneuploid clones as well as for coexisting diploid tumour cell populations in 14 cases. On the understanding that coexisting diploid and aneuploid tumour cell populations have a common clonal origin, these results provide evidence that aneuploid tumour clones typically develop from p53-deficient diploid progenitor cells. Loss of wild-type p53 function may therefore contribute to the development of aneuploidy in head and neck cancer.

Topics: Aneuploidy; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Clone Cells; Cohort Studies; Diploidy; Disease Progression; DNA, Neoplasm; Flow Cytometry; Genes, p53; Humans; Keratins; Mouth Neoplasms; Neoplasm Proteins; Neoplastic Stem Cells; Pharyngeal Neoplasms; Tumor Suppressor Protein p53

We studied three keratin (K) gene candidates, K13, K19, and K20 mRNAs, for detecting micrometastases in cervical lymph nodes (LNs) by reverse transcriptase-polymerase chain reaction (RT-PCR). Of 166 histologically metastasis-negative nodes, 24 micrometastatic LNs (14. 4%) were detected based on K13 gene expression. Keratin 19 mRNA is an inadequate marker for the genetic diagnosis due to not only illegitimate gene expression from lymphatic tissue but also gene expression from the ectopic salivary gland. Keratin 20 mRNA showed low sensitivity. It is suggested that K13 mRNA may be a promising tumor marker among these keratin genes for detecting the micrometastases in cervical LNs of oral cancer.

Topics: Gene Expression Regulation, Neoplastic; Humans; Keratins; Lymphatic Metastasis; Mouth Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured

To investigate whether the simple epithelial cytokeratin CK18 and CK19 can be used as a marker of oral precancerous lesions.. Formalin-fixed, paraffin-embedded tissue sections of normal oral mucosa, epithelial hyperplasia, mild epithelial dysplasia, moderate epithelial dysplasia, severe epithelial dysplasia and oral squamous cell carcinomas were stained with a CK18-specific antibody and CK19-specific antibody respectively by LSAB immunohistochemical method. The stained sections were observed under light microscopy. The results were described and analyzed with Rank Sum Test.. CK18 was not detected in normal and abnormal oral tissue sections. But in normal nonkeratinized mucosa, CK19 was detected in the basal cell layer dispersively. In epithelial dysplasia, CK19 was detected in the suprabasal cell layer and the number of CK19-positive cell layers was correlated with the dysplasia degree of epithelia. Furthermore, CK19 was detected in oral squamous cell carcinoma, especially in the poor-differentiated cancer cells.. CK19 expression in suprabasal cell layer of oral mucosa can be used as a candidate marker for diagnosis of oral precancerous lesions and determination of the differentiation level of oral squamous cell carcinoma.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Humans; In Vitro Techniques; Keratins; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions

The authors present an unusual case of an epithelial-myoepithelial carcinoma of the liver in a 67-year-old man who was admitted for resection of a gastric adenocarcinoma. At operation, a 3 x 3 cm mass in the right liver lobe was also removed. This mass consisted of duct-like structures with dual differentiation. The inner layer was composed of an epithelial lining, and the outer layer consisted of clear cells, all unrelated to the moderately well-differentiated gastric adenocarcinoma. The clear cells were positive for S-100 and alpha-smooth muscle actin, suggesting myoepithelial origin. The mass was considered to be low-grade epithelial-myoepithelial carcinoma. However, the patient had a history of an oral nodule present since childhood, resected 10 years previously. These slides were reviewed and revealed a mixture of clear cells and basal cells with squamous differentiation. In addition, there were duct-like structures with the two-layer pattern found in the liver tumor. This tumor had numerous mitotic figures and showed perineural invasion, suggesting a high grade of malignancy. These findings led to an interpretation of the oral tumor as also being epithelial-myoepithelial carcinoma, which had remained as "benign" for more than 50 years and subsequently underwent malignant transformation. During this long period, liver metastases may have occurred and remained low-grade. Alternatively, the liver and oral tumors may have arisen separately in the foregut during embryologic development, remaining low-grade until malignant transformation occurred.

Topics: Actins; Aged; Carcinoma; Humans; Immunoenzyme Techniques; Keratins; Liver Neoplasms; Male; Mouth Neoplasms; Palate; S100 Proteins

Mucosal oral squamous cell carcinoma (SCC) accounts for 3-5% of all reported cancers, with a 5-year survival rate of approximately 50%. Unfortunately, current detection means are of no value in diagnosing lesions early enough for cure, especially when they recur after resection. Postoperative radiotherapy and/or covering the resection site with reconstructive flaps (regional or free vascularized) often makes early diagnosis an impossible task.. The authors examined the detection and treatment monitoring capacity of two relatively new tumor markers in the serum of SCC patients, comparing their levels with those in patients with other oral/perioral malignancies or benign oral tumors and with disease free, posttreatment SCC patients and healthy controls.. Values of sensitivity, specificity, and positive and negative prediction for Cyfra 21-1 were 96%, 87%, 93%, and 53%, respectively, whereas those for tissue polypeptide specific antigen (TPS) were 69%, 87%, 93%, and 54%, respectively. Approximately 2-3 weeks after resection of the SCC lesion, Cyfra 21-1 and TPS levels were reduced by 47% (P < or = 0.003) and 36% (P < or = 0.041), respectively. Cyfra 21-1 levels in SCC patients were significantly greater than those of healthy patients by 73% (P < or = 0.0001), patients with benign tumors by 74% (P < or = 0.0003), and patients in disease remission by 66% (P < or = 0.0002). Similarly, the TPS levels of SCC patients were significantly greater than those of healthy patients by 59% (P < or = 0.0005), patients with benign tumors by 55% (P < or = 0.0001), and patients in disease remission by 59% (P < or = 0.0001). In two patients, a second, new SCC lesion was diagnosed within the follow-up period, with increased tumor markers noted concomitantly with the diagnosis.. The accumulated data point to the suitability of the clinical usage of these two markers, especially Cyfra 21-1, in the early detection of oral SCC lesions (primary, recurrent, or secondary) as well as for treatment monitoring. These results may open new avenues for the diagnosis and follow-up of these patients and hopefully improve their treatment outcome.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Humans; Jaw Neoplasms; Keratin-19; Keratins; Male; Middle Aged; Mouth Neoplasms; Peptides; Predictive Value of Tests; Sensitivity and Specificity; Time Factors; Treatment Outcome

Fas antigen is a cell-surface protein that transduces an apoptotic signal from the cell surface into the cytoplasm. The localization of Fas antigen in human oral squamous cell carcinoma (SCC) was examined by immunohistochemistry using a monospecific polyclonal antibody with a high titre. This antibody, designated as Fas D, was raised against a synthetic polypeptide segment corresponding to a specific extracellular domain of human Fas antigen (aa 104-114). Thirty-eight specimens of oral SCCs were stained with Fas D antibody and 26 (68%) reacted intensely. The specimens were graded as 'well', 'moderately', or 'poorly differentiated', according to the histopathological criteria. Out of 24 cases of the well differentiated tumours examined, 22 had reacted to Fas staining. The tumour cells formed nests that encompassed keratin pearls; staining was confined to cytoplasmic granules of peripheral cells. Among the moderately differentiated tumours, 4 out of 11 cases had reacted to Fas staining. No Fas-positive cells were observed in the poorly differentiated tumours, but only three specimens were examined. The expression of Fas antigen seems to be related to the degree of tumour differentiation.

Topics: Aged; Antibodies; Apoptosis; Carcinoma, Squamous Cell; Cell Differentiation; Cell Membrane; Coloring Agents; Cytoplasm; Cytoplasmic Granules; fas Receptor; Gene Expression Regulation, Neoplastic; Humans; Immunoenzyme Techniques; Immunoglobulin G; Immunohistochemistry; Keratins; Middle Aged; Mouth Neoplasms

We investigated the differences of Fourier transform infrared (FTIR) spectra between oral squamous cell carcinoma (OSCC) and normal gingival epithelium (NGE) or normal subgingival tissue (NST). We used 15 specimens of OSCC which had not been treated before measurement and 10 of NGE or NST. We also used cultured oral squamous cell carcinoma (COSCC) and the tissue (MSCC) which massed for 3 months after the cultured oral squamous cell carcinoma was transplanted into the lower back of a rat. Those tissue spectra were compared with the purified human collagens and human keratin. One half of every tissue specimen was measured with FTIR and the other half was investigated histologically. The differences of FTIR spectra between OSCC and NGE were observed in the bands between 1431 and 1482 cm(-1) and between 1183 and 1274 cm(-1). The shoulder at 1368 cm(-1) tended to disappear in OSCC, and the peaks at 1246 and 1083 cm(-1) found in NGE tended to shift to those at 1242 and 1086 cm(-1) in OSCC, respectively. The infrared spectrum of NST was noticed to be strongly influenced by the presence of collagen. Significant differences were also observed in the second derivative FTIR spectra between OSCC and NGE. Our data suggested that this infrared technique is applicable to clinical diagnostics.

Topics: Adolescent; Adult; Age Factors; Aged; Animals; Carcinoma, Squamous Cell; Cells, Cultured; Collagen; Epithelium; Female; Gingiva; Humans; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Rats; Reference Values; Sex Factors; Spectroscopy, Fourier Transform Infrared; Tumor Cells, Cultured

The expression of heat shock proteins HSP60 and HSP70 and cytokeratins CK1/10 and CK7/18 were compared in epithelium of oral lichen planus (OLP) lesions and oral fibromas using an avidin-biotin-peroxidase complex (ABC) immunohistochemical method. An immunostaining intensity distribution (IID) index was developed to assess staining intensity and the proportion of positively stained cells in different layers of the epithelium. The expression of HSP60 in the basal layer was significantly higher in OLP than in fibromas. No difference in HSP70 expression was evident between OLP and fibromas. The expression of CK1/10 in the epithelial basal and suprabasal layers was significantly higher in OLP than in fibromas. There was no demonstrable staining for CK7/18 in either OLP or fibromas. A significant correlation was evident between the expression of HSP60 and CK1/10 in the basal epithelial cells in OLP. The findings support a role for HSP60 in the pathogenesis of OLP. A unifying hypothesis of the pathogenesis of OLP, involving two sequential immune reactions, is proposed.

Topics: Adult; Basement Membrane; Chaperonin 60; Epithelial Cells; Fibroma; Genetic Predisposition to Disease; HSP70 Heat-Shock Proteins; Humans; Immunoenzyme Techniques; Keratins; Lichen Planus, Oral; Middle Aged; Mouth Neoplasms; Odds Ratio; Statistics, Nonparametric

The purpose of this study is to better estimate the true incidence of occult regional metastases associated with stage I and II squamous cell carcinoma of the oral cavity. The clinical and prognostic significance of micrometastatic disease discovered by cytokeratin immunoperoxidase reactivity in the previously pathologically N0 neck is also evaluated.. Forty patients treated between 1985 and 1996 with T1 or T2 squamous cell carcinoma of the lip and oral cavity were studied. All had primary surgical treatment including functional neck dissection. No metastases were demonstrated on hematoxylin and eosin microscopy. All specimens were reexamined with immunoperoxidase staining for cytokeratin.. Five percent of patients had micrometastatic disease. Retrospective analysis of patients with a minimum follow-up of 2 years has failed to show a statistically significant association between a positive cytokeratin analysis and poor locoregional control or overall survival.. Results suggest that the true incidence of occult metastases with carcinoma of the oral cavity is significantly higher than previously documented. However, the prognostic significance of these findings remains unclear.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Incidence; Keratins; Male; Middle Aged; Mouth Neoplasms; Neck Dissection; Neoplasm Staging; Prognosis; Retrospective Studies; Survival Analysis

Squamous cell carcinoma (SCC) of the head and neck is the sixth most frequent cancer worldwide. The survival rate is among the lowest of the major cancers and has not improved significantly in the past two decades. Extensive local invasion and regional lymph node metastasis are, in large part, responsible for the poor clinical outcome of these tumors. Keratin intermediate filaments are the most abundant cytoskeletal proteins in SCCs and regulate the migration of normal and transformed epithelial cells. Previous studies have shown that expression of the 40-kDa keratin K19 is dysregulated in SCCs arising from oral epithelium. Immunohistochemical experiments demonstrated that, while normal epithelium and dysplastic lesions expressed abundant K19 protein, invasive SCCs exhibited a patchy or negative staining pattern. We subsequently determined that K19 expression was consistently downregulated in seven SCC lines compared with normal epithelium. We therefore wanted to determine if K19 downregulation affected the invasive phenotype of these cells. We found that SCC lines which do not express K19 are significantly more invasive in vitro than those which retain expression of this gene. Stable expression of the K19 cDNA in K19 negative cell lines altered cell morphology and intercellular adhesiveness, and significantly decreased the number of cells able to migrate through a reconstituted basement membrane. Reduced invasiveness was not due to decreased metalloproteinase activity in the K19-expressing clones. We conclude that K19 overexpression in oral SCCs decreases their invasive potential by diminishing migratory capability.

Topics: Basement Membrane; Blotting, Western; Carcinoma, Squamous Cell; Cell Adhesion; Cell Line, Transformed; Cell Movement; Coloring Agents; DNA, Complementary; Down-Regulation; Electrophoresis, Polyacrylamide Gel; Epithelial Cells; Fluorescent Antibody Technique; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Lymphatic Metastasis; Metalloendopeptidases; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness; Prognosis; Survival Rate; Tumor Cells, Cultured

Cytokeratin (CK) alterations have been reported in carcinomas from different anatomical sites, and these have been associated with specific aspects of tumour behaviour. In order to assess the relationships between CK modifications and future tumour behaviour, we conducted the present prospective study on 26 squamous cell carcinomas (SCC) of oral and pharyngeal mucosae and corresponding controls. Cytokeratins were investigated using two-dimensional gel electrophoresis and immunofluorescence techniques. All healthy tissues, oral lining and oropharyngeal mucosae, expressed the oesophageal type CKs, including CK 19. Other simple epithelial CKs (7, 8, 17 and 18) were not detected. In carcinomas originating from corresponding sites, expression of oesophageal CKs varied widely from one specimen to another, and simple epithelial keratins were often found. Statistical analysis indicated correlations between CK expression and the clinicopathological data of SCC patients. Small tumour size was strongly associated with the expression of CKs 10 and 19. Interestingly, an absence of lymph node involvement was significantly associated with CK 18 expression. Tumours giving rise to recurrences, metachronous tumours, and distant metastasis were significantly associated with an absence of CK 13. These results suggest that CKs 10, 19, 18 and 13 could be reliable diagnostic and prognostic markers in the assessment of oral and pharyngeal squamous carcinomas.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Chi-Square Distribution; Disease Progression; Electrophoresis, Gel, Two-Dimensional; Female; Fluorescent Antibody Technique, Indirect; Humans; Keratins; Male; Middle Aged; Molecular Weight; Mouth Mucosa; Mouth Neoplasms; Neoplasm Metastasis; Neoplasm Proteins; Neoplasm Recurrence, Local; Pharyngeal Neoplasms; Prognosis; Prospective Studies

Immunohistochemistry with monospecific antibodies was used to study the expression patterns of cytokeratins (Cks) and vimentin in non-dysplastic lesions of the oral cavity, including lichen planus and fibromas. In hyperplastic lesions, Ck expression did not deviate significantly from the normal non-keratinizing squamous epithelium of the oral cavity. Hyperkeratotic lesions showed pronounced aberrations in their Ck profile. These lesions were characterized by extended expression of the keratinization marker Ck 10, the basal cell Ck 14 and the hyperproliferation-associated Ck 16 in the suprabasal compartment. The stratification markers Cks 4 and 13 showed a decreased expression. Coexpression of Cks and vimentin was found in lesions having accumulations of inflammatory cells in the subepithelial cell layer. These changes are felt to characterize benign mucosal lesions without dysplasia and might be helpful for distinguishing these lesions from potentially malignant ones.

Topics: Antibodies, Monoclonal; Cell Division; Epithelial Cells; Epithelium; Fibroma; Gene Expression Regulation; Humans; Hyperplasia; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Leukoplakia, Oral; Lichen Planus, Oral; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Vimentin

The bcl-2 proto-oncogene is a known inhibitor of apoptosis; in normal human stratified squamous epithelium, its expression is restricted to the basal cell layer. To investigate the functional role of bcl-2 protein in the process of differentiation of oral keratinocytes, bcl-2 expression vector was transfected into SCC-25 cells, which normally undergo squamous cell differentiation in vitro while expressing specific differentiation markers, e.g., keratin 10/11 and involucrin. In bcl-2 transfected SCC-25 cells, the expression of these differentiation markers was markedly suppressed. The bcl-2 proto-oncogene may play a critical role in opposing the commitment to terminal differentiation and apoptosis of oral keratinocytes.

Topics: Apoptosis; Carcinoma, Squamous Cell; Cell Differentiation; DNA Fragmentation; Down-Regulation; Genes, bcl-2; Humans; Immunoenzyme Techniques; Keratinocytes; Keratins; Mouth Neoplasms; Protein Precursors; Proto-Oncogene Mas; Proto-Oncogene Proteins c-bcl-2; Tumor Cells, Cultured

The effects of an induction chemotherapy with THP-adriamycin, cisplatin, and peplomycin (TPP) were studied in 32 patients with operable oral cancer. The histological evaluation according to the Shimozato-Oboshi classification was Grade (G) IV in ten cases (31.3%), GIII in one case, and GIIb in four cases. Induction of apoptosis and differentiation-inducing effects, hyperkeratinization or bone formation, were observed in some cases. The overall clinical response rate and histological response rate were 63% and 47%, respectively. Grade III was obtained in seven metastatic lymph nodes of three patients. The expressions of PCNA, p53, and AgNORs before and after chemotherapy were studied. The prechemotherapeutic PCNA positive cell index (PI) of the highly responsive tumors (GIII, IV) was significantly lower than that of the poorly responsive tumors (G0-IIb) (P < 0.01). Similar results were obtained in the evaluation of p53 PI (P < 0.05), suggesting that PCNA and p53 are useful biomarkers for predicting the efficacy of TPP chemotherapy.

Topics: Antibiotics, Antineoplastic; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Cisplatin; Coloring Agents; Doxorubicin; Female; Forecasting; Gene Expression Regulation, Neoplastic; Humans; Keratins; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Nucleolus Organizer Region; Osteogenesis; Peplomycin; Proliferating Cell Nuclear Antigen; Remission Induction; Silver; Treatment Outcome; Tumor Suppressor Protein p53

Precise regulation of cell organization results in well-differentiated tissue structures and continuous renewal of the oral epithelium maintaining a highly ordered tissue architecture. Here we demonstrate that FT-IR microspectroscopy, performed on sections of cancerous tissue biopsies, is capable of generating biochemical maps that show the distribution and any abnormal concentration of individual classes of biomolecules. Oral epithelia affected by cancer (squamous cell carcinoma) show many abnormal changes in morphology, of which the formation of keratin pearls is only one. Spectra from selected pearl areas demonstrate that these structures contain not only abnormal keratin concentrations but also seem to be stabilized by surrounding collagen fibers. Infrared image maps reveal that in the center of keratin pearls the concentration of protein (cytokeratins) is abnormally high, that DNA is absent and that the cell membrane fluidity is reduced. This suggests that cells are structurally destroyed and transformed into nuclei-free horny cells, simulating normal differentiation and epithelial growth. We also introduce a new analysis modality, two-dimensional (2D) tissue classification, and apply it to establish spectral similarities between different tissue structures. A total of 315 spectra, recorded for the original map, were analyzed by pattern recognition methods, classified and re-assembled into new maps based on their spectral similarities. The re-assembled maps clearly indicate significant tissue changes outside the pearls, suggesting early biochemical changes that accompany abnormal growth. Employing this 2D analysis modality in combination with infrared histopathology may be relevant to tumor diagnosis and prognosis.

Topics: Cluster Analysis; Humans; Keratins; Mouth Neoplasms; Neoplasms, Squamous Cell; Spectroscopy, Fourier Transform Infrared

Serologic identification of the fragment of cytokeratin 19 known as CYFRA 21-1 has been used for early detection of squamous cell carcinoma of the lung. The sensitivity of the CYFRA 21-1 assay in detecting oral cancers is lower than that in detecting lung cancers. To clarify the reason for this, we compared the cytokeratin expression in these cancers, with special reference to cytokeratin 19.. Oral squamous cell carcinomas and lung squamous cell carcinomas were immunostained with cytokeratin 19, cytokeratin 10, and cytokeratin 13 antibodies. Staining intensity was scored on a graduated scale from 0 to 4.. With respect to cytokeratin 19, the stainings of all lung cancers were scored as 4, which indicates a greater expression of cytokeratin 19 than is seen in oral cancers (p < 0.01). With an average cytokeratin 19 staining score of 1.67, oral cancers ranked lowest among the antibodies. Squamous cell carcinomas of the maxillary sinus arising from pseudostratified ciliated epithelium were highly expressive of cytokeratin 19. A marker for keratinizing cells (cytokeratin 10) and a marker for squamous cells (cytokeratin 13) were expressed more frequently and intensely in oral cancers (p < 0.01) than in lung cancers (p = 0.019).. From the viewpoint of immunohistochemistry, cytokeratin 19 was found to be a tumor marker with low specificity and sensitivity in oral cancers. The staining results suggested that poor expression of cytokeratin 19 by oral squamous cell carcinoma may result in a low serum value of CYFRA 21-1 in patients with this condition.

Topics: Antibodies; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Coloring Agents; Epithelial Cells; Epithelium; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratin-19; Keratins; Lung Neoplasms; Maxillary Sinus Neoplasms; Mouth Neoplasms; Sensitivity and Specificity

Reduction or loss of the intercellular junctions known as desmosomes may contribute to the invasive and metastatic behaviour of various carcinomas. Previous studies have shown that metastasis of oral squamous cell carcinomas of the head and neck correlates with a reduction in immunohistochemical staining for desmoplakin and desmoglein at the invasion front. The primary aim of the present study was to extend these observations to include a third component of desmosomes, the glycoprotein desmocollin. An additional aim was to determine whether the differentiation status of tumours is reflected in their staining for cytokeratins 1, 13, and 19, and, if so, whether these parameters correlate with desmosomal staining and/or metastasis. The study included 54 primary tumours of which 28 showed lymph node metastases. The results of this investigation show that tumours can be divided into three groups according to whether they have lost staining for no, one or more than one desmosomal component. A statistically significant correlation was found between the number of desmosomal components lost and metastasis. Tumours could also be divided into five groups according to their staining for different combinations of cytokeratins. Furthermore, differentiation status as indicated both histologically and by cytokeratin staining correlated with reduced desmosomal staining and metastasis. Tumours were also examined for intensity of staining for the adhesion molecule E-cadherin. Reduction in E-cadherin staining was correlated with mode of invasion and with reduction in desmosomal staining, but not with poor differentiation as indicated by cytokeratin staining. The results of this extensive study reinforce the view that adhesive junctions and adhesion molecules contribute to the suppression of tumour invasion and metastasis.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cadherins; Carcinoma, Squamous Cell; Cell Differentiation; Cytoskeletal Proteins; Desmocollins; Desmogleins; Desmoplakins; Desmosomes; Humans; Immunoenzyme Techniques; Keratins; Lymphatic Metastasis; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Proteins

This study aimed to ascertain whether cathepsin D expression could be related to the stage of differentiation of oral tumors.. Human oral biopsies of 10 squamous cell carcinomas and of the corresponding perilesional normal tissues were used. The tumors had all been clinically graded as advanced stage but nonmetastatic; five were classified histopathologically as poorly differentiated.. The gene expression of cathepsin D and keratin K13 in the biopsies was measured by reverse transcription polymerase chain reaction. Ratios of tumor-to-control readings helped compensate for sample variability.. Keratin K13, as a suprabasal cell marker, tended to confirm the histological grading of the tumors (but was not otherwise useful in distinguishing tumors from normal tissue). Substantial overexpression of cathepsin D was found in the poorly differentiated tumors.. Cathepsin D overexpression is considered a prognostic indicator of metastasis. In this sample, it was also associated with dedifferentiation. Cathepsin D might serve as a valuable gauge in clinical exploration of the connection between dedifferentiation and metastasis.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cathepsin D; Cell Transformation, Neoplastic; Female; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Polymerase Chain Reaction; Prognosis; RNA-Directed DNA Polymerase; RNA, Messenger

Fourier transform infrared (FTIR) microspectroscopy is emerging as a promising new tool for histopathological investigations of tissue histochemistry. This study was designed to assess whether changes in tissue biochemistry induced by well-differentiated and poorly differentiated oral/oropharyngeal squamous cell carcinoma (SCC) can be detected by infrared spectroscopy. The biopsies analyzed were each proven SCC positive and compared with tissue taken from the contralateral normal site. Individual infrared spectra, recorded from specific tissue areas, were correlated with histopathological structures normally found in the oral mucosa. Infrared mapping of these areas allows the generation of biochemical images of molecular structures such as lipids, sugars, and proteins. The visualization of DNA and tissue structures containing keratin (well expressed in all epithelia) reveals distinct differences between normal and SCC-positive biopsies. Bivariate histogram analysis of cell components (e.g., DNA and keratin) indicated that cancer cells produce a relatively homogeneous and clearly abnormal cell biochemistry, whereas differentiated epithelial cells present a very heterogeneous distribution of cellular components. Using these features, tissue containing abnormal or cancer cells can easily be distinguished from normal epithelial structures. The abnormal keratin distribution in poorly differentiated SCC and in keratin pearls (present only in well-differentiated SCC) offers insight into the process of malignant tissue transformation in squamous epithelium. Applying infrared microspectroscopy in combination with bivariate statistics to histopathological tissue thin sections provides a potential diagnostic tool for detection of cell changes in epithelial cancers.

Topics: Aged; Aged, 80 and over; Carcinoma, Squamous Cell; DNA; Female; Humans; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Pharyngeal Neoplasms; Spectroscopy, Fourier Transform Infrared

Cytokeratin (CK) expression was studied in buccal mucosa (BM) from 20 leucoplakia and 7 submucous fibrosis patients using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting and two-dimensional gel electrophoresis with iso-electric focussing (IEF) as the first dimension. Normal BM expresses CK 4, 5, 13, 14 and perhaps 19. Of 20 leucoplakia samples analysed, CK 5 was not detected in 17 samples, while CK 14 was not found in 13 samples. CK 1 and CK 8 were aberrantly expressed in six and seven samples, respectively. CK expression in contralaterally collected uninvolved tissues from 3 patients showed a normal pattern in two samples. Non-expression of CK 5 was observed in five of seven submucous fibrosis samples, while CK 14 was not detected in only two samples. CK 8 was aberrantly expressed in three samples. All the leucoplakia patients were chronic tobacco chewers. Thus, non-expression of CK 5 may be an early event occurring in tobacco-associated pathological changes in the BM.

Topics: Biomarkers, Tumor; Electrophoresis, Polyacrylamide Gel; Humans; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Oral Submucous Fibrosis; Plants, Toxic; Precancerous Conditions; Tobacco, Smokeless

In stratified squamous epithelia, altered expression of keratins (Ks) is one possible marker of malignant potential. In the epithelium of the uterine cervix, presence of human papillomaviruses (HPVs) is increasingly regarded as a marker of risk for cervical cancer. However, a similar role in oral cancer and precancer remains controversial. To address these questions, formalin-fixed, paraffin-embedded oral carcinomas from Sudanese snuff dippers (n=14) and oral carcinomas from Sudanese (n=14), Swedish (n=19) and Norwegian (n=41) non-snuff dippers were examined by immunohistochemistry for expression of K types 13, 14 and 19 using monoclonal antibodies. HPV infection was searched for in all the carcinomas by in situ hybridization (ISH) using the cocktail HPV OmniProbe and the ViraType probe. Carcinomas from Sudanese (snuff dippers/non-snuff dippers) were also examined for HPV infection by polymerase chain reaction (PCR) using the general HPV primers GP5+/GP6+. For the oral carcinomas from snuff dippers, moderate to intense expression of K13 (71%; 10/14), K14 (86%; 12/14) and K19 (93%; 13/14) was found. For the oral carcinomas from non-snuff dippers, weak to moderate expression of K13 (64%; 47/74), K14 (43%; 32/74) and K19 (45%; 33/74) was found. HPV DNA was not detected in any of the carcinomas from three countries when examined by ISH. The Sudanese (from snuff dippers/non-snuff dippers) oral carcinomas were also negative for HPV DNA with the PCR. The present study shows that (i) there is a high level of expression of K13, K14 and K19 in oral carcinomas from snuff dippers compared to those from non-snuff dippers, (ii) this high level of expression may arise from dysregulation of keratinocyte proliferation and maturation caused by damaging effects of snuff, (iii) the HPV genome is not found in Sudanese (snuff dippers/non-snuff dippers), Swedish or Norwegian oral carcinomas, and (iv) this may suggest that these viruses do not play a prominent role in the aetiology of oral carcinomas from these countries.

Topics: Carcinoma, Squamous Cell; Female; Humans; In Situ Hybridization; Keratin-14; Keratins; Male; Mouth Neoplasms; Norway; Papillomaviridae; Papillomavirus Infections; Plants, Toxic; Polymerase Chain Reaction; Sudan; Sweden; Tobacco, Smokeless; Tumor Virus Infections

P53 is overexpressed in more than 50% of all human cancers. A previous study suggested that p53 was also overexpressed in oral papillomas. This study was carried out to investigate whether p53 expression was correlated with expression of the cellular proliferation marker Ki-67 and the epithelial differentiation marker cytokeratin-4 (CK4) in oral papillomas. Formalin-fixed paraffin-embedded sections of 30 oral papilloma specimens and 30 unmatched normal oral mucosal specimens were processed for immunohistochemistry, using an avidin-biotin-peroxidase procedure and monoclonal antibodies. A semiquantification analysis on p53 and Ki-67 labeling indices was performed. Twenty-eight of 30 (93%) papilloma specimens were positive for p53. The percentage of p53-positive cells in the basal layer was 60.4 +/- 14.8 (mean +/- SD, n = 28), and that of Ki-67-positive cells was 26.7 +/- 14.4. There was no correlation between expression of p53 and that of Ki-67. Expression of CK4 was inversely correlated with the expression of Ki-67 but not correlated with the expression of p53.

Topics: Antigens, Viral, Tumor; Biomarkers, Tumor; Gene Expression Regulation, Neoplastic; Humans; Immunoenzyme Techniques; Keratins; Ki-67 Antigen; Mouth Neoplasms; Papilloma; Tumor Suppressor Protein p53

The granular cell tumour (myoblastoma, Abrikosoff's tumour) and the congenital epulis in newborns (Neumann tumour) are two lesions rarely found in the oral cavity, whose histogenetic origin is highly controversial. This work analyses using immunohistochemical techniques 15 cases of myoblastomas and two of congenital epulis with different mono- and poly-clonal antibodies. Positive immunostaining was found for S-100 protein and neuron-specific enolase in all the cases of myoblastoma, and for vimentin and carcinoembryonic antigen in some cases. No immunoreactivity was observed for any of the other 13 antibodies used in congenital epulis.

Topics: Actins; Adolescent; Adult; Carcinoembryonic Antigen; Coloring Agents; Desmin; Female; Gingival Neoplasms; Granular Cell Tumor; Humans; Immunohistochemistry; Infant, Newborn; Keratins; Male; Mouth Neoplasms; Mucin-1; Myelin Basic Protein; Neoplasms, Muscle Tissue; Neurofilament Proteins; Phosphopyruvate Hydratase; S100 Proteins; Vimentin

To characterize further the nature of calcifying odontogenic cyst (COC), we studied histologically and immunohistochemically an extraosseous and two intraosseous lesions. The extraosseous COC was in continuity with the stratified squamous epithelium of the alveolar mucosa. Immunostaining with monoclonal antibodies showed reactivity of both low- and high-molecular-weight cytokeratins, the degree of coexpression decreasing with the increasing morphological diversity of the cyst/tumour epithelium. Staining for the matrix glycoprotein tenascin-C was seen not only in the connective tissue, where its distribution patterns corresponded to the stage of hard tissue formation, but also in epithelial elements. The staining patterns were analogous to those described during normal tooth formation. Both the morphological characteristics and expression patterns of the various cytokeratin types and tenascin-C implied that COC represents a pathological counterpart of normal odontogenesis. In the case of the extraosseous COC, the correspondence could be traced back to early stages of tooth development.

Topics: Adolescent; Adult; Aged; Antibodies, Monoclonal; Cell Lineage; Child; Coloring Agents; Connective Tissue; Epithelium; Female; Gene Expression Regulation, Neoplastic; Gingiva; Glycoproteins; Humans; Immunohistochemistry; Keratins; Male; Mandibular Neoplasms; Mouth Mucosa; Mouth Neoplasms; Odontogenesis; Odontogenic Cyst, Calcifying; Radicular Cyst; Tenascin

The expression of the adhesion molecule CD44 variant 6 (CD44v6) was studied immunohistochemically on 38 oral squamous cell carcinomas (SCCs) and 10 biopsies of healthy oral mucosa. The relationship between the expression of CD44v6 and regional lymph node metastasis was also investigated. The expression of CD44v6 was apparently down-regulated in oral SCC, but not in normal oral mucosa. Carcinomas expressing lower levels of CD44v6 exhibited more frequent regional lymph node metastasis. The expression of CD44v6 showed no statistically significant relationship to the degree of differentiation, but tended to be down-regulated in poorly differentiated carcinoma. No significant relation was found between the expression of CD44v6 in primary and metastatic lesions.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Cell Differentiation; Cell Membrane; Coloring Agents; Down-Regulation; Epithelium; Exons; Fibroblasts; Gene Expression Regulation, Neoplastic; Humans; Hyaluronan Receptors; Immunoenzyme Techniques; Keratins; Lymphatic Metastasis; Mouth Floor; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness; Tongue Neoplasms

This study evaluated use of a combination assay of tumor markers in the diagnosis of oral squamous cell carcinoma.. Serum levels of four tumor markers (carcinoembryonic antigen [CEA], squamous cell carcinoma antigen [SCCA], immunosuppressive acidic protein [IAP], and cytokeratin 19 fragment [Cyfra]) were simultaneously measured in 42 patients with oral squamous cell carcinoma (O-SCC) and in 12 patients with oral benign diseases.. The positive rates were 31.0% for CEA, 38.1% for SCCA, 52.4% for IAP, and 38.1% for Cyfra in patients with O-SCC. These rates were significantly different (P < .01) from those of control patients with oral benign diseases. The sensitivity (81.0%) and accuracy (77.8%) of the combination assay uses higher than that obtained with individual markers.. A combination assay with CEA, SCCA, IAP, and Cyfra may be useful for the screening of patients with suspected oral squamous cell carcinoma.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Chi-Square Distribution; Female; Humans; Keratins; Male; Middle Aged; Mouth Diseases; Mouth Neoplasms; Neoplasm Proteins; Sensitivity and Specificity; Serpins

Topics: Adult; Biopsy; Connective Tissue; Diagnosis, Differential; Epithelium; Female; Follow-Up Studies; Humans; Keratins; Melanocytes; Mouth Neoplasms; Neoplasms, Glandular and Epithelial

In a clinical trial the effect of chemoprevention with beta-carotene, vitamin E and C on dysplastic tissue was studied. The study included 24 patients with oral leukoplakia and 24 patients after radical resection of a primary oral cancer. There was a reduction of increased cell kinetic parameters like the S-phase portion or the average number of nuclear-organizer regions (NOR) per cell nucleus, a decrease of the micronuclei portion and a normalization of the cytokeratin gene-expression. The general response was 97.5%. Stopping the alcohol and tobacco abuse the effect of the antioxidative vitamins on redifferentiation of the oral mucosa was more intense than by persistance of the alcohol and tobacco abuse, but a long term prevention seems to be ineffective.

Topics: Antioxidants; Ascorbic Acid; beta Carotene; Biopsy; Cell Differentiation; Cohort Studies; Drug Therapy, Combination; Humans; Keratins; Leukoplakia; Mouth Mucosa; Mouth Neoplasms; Nucleolus Organizer Region; S Phase; Time Factors; Vitamin E

Since it was first described in 1986, basaloid-squamous cell carcinoma (BSC) has been considered a distinct variant of squamous cell carcinoma that occurs in a variety of anatomic sites, including the head and neck region. We report the characterization of the first cell line established from a basaloid-squamous cell carcinoma of the floor of the mouth. The cell line exhibited a highly invasive capacity, indicating that BSC has very aggressive behavior. This cell line may be a useful model for elucidation of the biological characteristics of BSC.

Topics: Actins; Biology; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cell Division; Cell Movement; Cell Separation; Culture Media; Endothelium; Epithelial Cells; Fibroblasts; Humans; Keratins; Male; Middle Aged; Mouth Floor; Mouth Neoplasms; Mucin-1; Neoplasm Invasiveness; S100 Proteins; Tongue Neoplasms; Tumor Cells, Cultured

The single cell gel electrophoresis (SCGE) assay, also known as the comet assay, is a cytogenetic technique for measuring and analyzing DNA single stranded breaks (SSB) and/or alkali labile sites within individual cells. Peripheral blood leukocytes of 22 oral squamous cell carcinoma (OSCC) patients were subjected to SCGE and the DNA damage levels (SSB) were quantified with respect to clinical staging and histopathologic grading. Highly statistically significant differences in DNA damage levels were found between normal subjects and patients with OSCC of the same age group. DNA damage levels were altered in all clinical stages and histopathological grades of oral squamous cell carcinoma. The differences were generally significant between all the clinical stages of OSCC, while in histopathologic grading the results were significant only between grades I and III. The results support the concept of a systemic host response in malignancy.

Topics: Adult; Age Factors; Alkalies; Blood; Carcinoma, Squamous Cell; Case-Control Studies; Cell Nucleus; Cytogenetics; DNA Damage; DNA, Neoplasm; DNA, Single-Stranded; Electrophoresis, Agar Gel; Female; Humans; Keratins; Leukocytes; Male; Middle Aged; Mitosis; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Staging; Plasma Cells

Cytokeratin (CK) expression in untreated, paraffin-treated or dimethylbenzanthracene (DMBA)-treated hamster cheek pouch epithelium was investigated utilizing monoclonal antibodies AE1 or AE3, which react with type I or type II CKs, respectively, and by in situ hybridization utilizing type I CK-specific probes. The latter were isolated from a cDNA library of hamster cheek pouch mRNA and designated CK 13 and CK 10 based on their respective homologies (> 95% amino acids) with murine CK 13 and human CK 10. Treatment of hamster cheek pouch epithelium with DMBA resulted in increased expression of type I CK, detected immunohistochemically with monoclonal AE1, but decreased expression of type II CKs detected with AE3. Despite an overall increase in type I CKs, in situ hybridization demonstrated differential expression of type I CKs with altered distribution of CK 13 mRNA and reduced expression of CK 10 mRNA, providing additional sensitive markers for DMBA-associated changes in CKs. These changes were constant at 2 to 22 weeks in the pre-neoplastic and neoplastic epithelium following the initial application of DMBA.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinogens; Cricetinae; DNA, Complementary; Epithelium; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Male; Mice; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; RNA, Messenger; Sequence Homology, Amino Acid

The carcinogen 7,12-dimethylbenz(a)anthracene (DMBA) has been used to induce oral carcinogenesis of the hamster buccal mucosa in an experimental model that exhibits many of the genetic, biochemical and pathological features of human oral squamous cell carcinoma. To complement this in vivo process we have established an in vitro transformation procedure that involved the treatment of normal hamster oral mucosal keratinocytes (NHKs) with DMBA. Uptake of DMBA in NHKs was verified by observing autofluorescence of DMBA in the oral mucosal cells. Treatment doses ranged from 5, 50 and 200 ng and the NHKs were generally treated with DMBA for 1-14 days. The 200 ng dose proved to be toxic to these cells. The 5 and 50 ng treatments were found to maintain the viability of the NHKs and demonstrate anchorage-independent agarose growth, producing 18 and 40 colonies, respectively, after 14 days of treatment. Characterisation assays included determinations for cellular growth through plating efficiency, counting of cell colony number, and 3H-thymidine incorporation. Differentiation was ascertained by counting cornified cells, specification of either high or low molecular weight keratins, the percentage of cells expressing gamma glutamyl-transpeptidase (GGT), the level of p53 expression, and a determination of cell cycle. After 24 h the plating efficiency of the NHKs was found to be slightly increased following treatment with a 5 or 50 ng dose of DMBA compared to the untreated NHKs. After 14 days of incubation these doses also enhanced the number of colonies formed by the NHKs (e.g. plating efficiency). In contrast, the number of cornified cells was reduced in these colonies, while immunohistochemistry disclosed an increase in the number of NHKs expressing low molecular weight keratins, significantly lower levels of high molecular weight keratins and high levels for GGT. Flow cytometric analysis verified an increase in p53 expression (e.g. p53 wild type, 19% and p53 mutant, 66%). Cell cycle analysis of NHKs treated with DMBA (5 ng) demonstrated a shift in the number of cells in S phase (17.2%) and G2 + mitosis (11.0%). Cells from this DMBA treatment group were injected into syngeneic hamster recipient buccal pouches (10 x 10(6) cells/0.25 ml). Squamous carcinomas grew in four of six hamster buccal pouches as determined by histopathological analysis. The in vitro assay system will enhance our ability to define the genetic and molecular changes related to chemical carcinogen

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinogens; Carcinoma, Squamous Cell; Cell Culture Techniques; Cell Cycle; Cell Transformation, Neoplastic; Cheek; Cricetinae; Dose-Response Relationship, Drug; Keratinocytes; Keratins; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Neoplasm Transplantation

To test and optimize photodynamic therapy of early cancers in the upper aero-digestive tract and oesophagus, we sought an appropriate animal model, which was found in the 7,12-dimethylbenz[a]anthracene-induced early squamous cell carcinoma in the Golden Syrian hamster. This chemically induced neoplasm is shown, by histology and immunohistochemistry, to pass through similar stages of early cancer development as its human counterpart. Its time sequence is highly reproducible, leading to a well differentiated carcinoma in situ and microinvasive carcinoma in the hamster cheek pouch over a period of 10 weeks.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinogens; Carcinoma in Situ; Carcinoma, Squamous Cell; Cheek; Cricetinae; Disease Models, Animal; Esophageal Neoplasms; Keratins; Male; Mesocricetus; Mouth Neoplasms; Neoplasm Invasiveness; Photochemotherapy; Precancerous Conditions

Cytokeratin (CK) expression was studied in squamous cell carcinomas of different subsites in the head and neck by using cryostat sections from 27 head and neck squamous cell carcinomas (HNSCCs) and 6 cell lines established from HNSCC. All tissues were analyzed immunohistochemically with a panel of monospecific anti-keratin monoclonal antibodies. Most carcinomas recapitulated the expression pattern of keratins present in the basal layer of normal epithelium from the site of tumor origin. Regional differences in the expression of simple-epithelial type of keratins in stratified (pseudostratified) epithelia were to a large extent repeated in corresponding carcinomas. In the present study, localization of various keratins were surveyed and CK 18 specific monoclonal antibodies were specifically used to distinguish SCCs of the larynx or hypopharynx from SCCs of the oral cavity. CK 18 staining of almost all tumor cells was detected in 11 of 12 SCCs of the larynx and hypopharynx, but was only detected sporadically in 3 of 9 SCCs of the oral cavity. The present results show that CK 18 typing might be useful for distinguishing sites of origin of various HNSCCs. Findings also indicate that CK 18 expression in SCC might be modulated by microenvironmental factors.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Cell Line; Diagnosis, Differential; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Hypopharyngeal Neoplasms; Hypopharynx; Immunoenzyme Techniques; Keratins; Laryngeal Neoplasms; Larynx; Mouth Mucosa; Mouth Neoplasms

Expression of human leukocyte elastase inhibitor, elafin, otherwise known as skin-derived antileukoproteinase inhibitor (SKALP), was investigated in normal and abnormal oral tissues using a specific anti-SKALP rabbit antiserum. Weak staining was observed in keratinizing epithelia of normal oral mucosa but not in non-keratinizing mucosa. Increased expression was also observed in the suprabasal layers of dysplastic oral epithelia and in well-differentiated squamous cell carcinoma, but not in basal cell carcinoma. A uniform strong expression was observed in all supra-basal layers of odontogenic keratocyst epithelia, except in regions where inflammatory infiltrate was adjacent to keratocyst epithelia. In contrast, elafin expression in a small number of dentigerous cysts and ameloblastomas was more patchy. The increased levels of elafin in keratocyst epithelia and dysplastic tissue may be a cellular homoeostatic response to generate a protective barrier preventing proteolytic degradation of underlying elastic tissue.

Topics: Ameloblastoma; Animals; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Dental Sac; Dentigerous Cyst; Elastic Tissue; Epithelium; Gene Expression Regulation; Gene Expression Regulation, Neoplastic; Gingiva; Homeostasis; Humans; Keratins; Mouth Mucosa; Mouth Neoplasms; Odontogenic Cysts; Peptide Hydrolases; Proteinase Inhibitory Proteins, Secretory; Proteins; Rabbits; Serine Proteinase Inhibitors

We investigated the incidence of micrometastases from squamous cell carcinomas of the head and neck in neck dissection specimens originally staged as pNO. A total of 76 dissection specimens from 60 patients were evaluated using serial microscopic sectioning in 10-microns intervals, H & E staining, and immunostaining with an antibody to pan-cytokeratin. Examination of 1,020 lymph nodes from 76 neck dissection specimens revealed 8 micrometastases (7.9%) in 6 specimens from 6 patients with oral and pharyngeal primaries, resulting in upstaging. Six micrometastases were located in lymph nodes 3 to 6 mm in diameter. The surgeon should be aware of the relatively high incidence of micrometastases from oral and pharyngeal carcinomas, which are undetectable preoperatively or by routine histopathologic examination. Primary tumor site (oral cavity and pharynx) and certain features of the primary can delineate a group of patients with a higher risk of harboring occult metastases who may benefit from elective treatment of the neck.

Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Histological Techniques; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Lymph Node Excision; Lymphatic Metastasis; Mouth Neoplasms; Neoplasm Staging; Pharyngeal Neoplasms; Prognosis; Retrospective Studies

Specific mRNA and protein for two major keratins, K14 and K19, were investigated in normal, dysplastic and malignant oral epithelia by combined in situ hybridization and immunohistochemistry. In normal epithelia, K14 mRNA and protein were present almost exclusively in the basal layer of non-cornified, and in rete-processes of cornified, sites. Dysplastic epithelium showed irregular extension of the K14 transcript and protein into superficial cells. In squamous cell carcinoma (SCC), K14 transcript was abundant in most samples whilst in one poorly differentiated carcinoma mRNA but no protein was detected. K19 mRNA and its protein were present predominantly in basal cells of noncornified epithelium, whereas in cornified epithelium only mRNA was detected. In dysplasias, K19 transcript was detected in all specimens but its protein was absent in most cases. Even more variations of K19 expression were observed in SSC. These findings indicate differences in the control of expression of K14 and K19 in normal epithelia and show that regulation is further disturbed during dysplastic change and malignancy.

Topics: Autoradiography; Carcinoma, Squamous Cell; Epithelium; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Neoplasm Proteins; RNA Probes; RNA, Messenger

The aim of the present study was to investigate the sequential expression of placental glutathione S-transferase (GST-P) during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch squamous cell carcinogenesis. Both immunohistochemical and immunoblot analyses were employed to detect the epithelial GST-P in hamster buccal pouch mucosa over a 15-week treatment regimen. No GST-P positivity was demonstrated in the pouches of the control group. GST-P positive cells were first noted as early as 1 week after DMBA applications. A gradual increase in both the mean number and size of GST-P-positive foci was noted in the first 12 experimental weeks, but a plateau level was approached thereafter. The early GST-P-positive-area were located in the basal layer, or occasionally in the middle layer, of DMBA-treated hamster buccal pouch mucosa. Later, the stained sites became enlarged and were scattered randomly in different layers or in the whole thickness of the dysplastic and non-dysplastic epithelium. The keratin layer was only occasionally involved during the first 12 weeks of DMBA treatment but positive staining was more noticeable in the final stage of the experiment. Both exophytic (8-12 weeks) and invasive (13-15 weeks) squamous cell carcinomas showed GST-P positivity, in both cytoplasmic and nuclear components. Immunoblot analysis revealed no band in the crude tissue extracts of the control pouches whereas GST-P polypeptide of molecular weight approximately 26 kD was demonstrated in DMBA-treated pouches over the whole 15-week treatment regimen. Results of the present work indicate that GST-P is a stable and persistent label for almost all of the carcinogen-altered cells during DMBA-induced hamster buccal pouch carcinogenesis. Immunohistochemically detectable GST-P may be a potential marker throughout oral chemical carcinogenesis.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Biomarkers, Tumor; Carcinogens; Carcinoma, Squamous Cell; Cell Nucleus; Coloring Agents; Cricetinae; Cytoplasm; Epithelium; Gene Expression Regulation, Neoplastic; Glutathione Transferase; Immunoblotting; Immunohistochemistry; Keratins; Male; Mesocricetus; Molecular Weight; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness; Placenta; Time Factors

It is generally agreed that there is a need for a routine, non-invasive screening procedure for oral cancer particularly of high risk groups. Refinements in oral exfoliative cytology now make this technique worthy of consideration for such screening. This study assesses the utility of monitoring cytokeratin expression in smears of oral cancer in comparison with assessing the keratin expression in corresponding biopsies. Smears and biopsies were taken from 34 patients with oral cancer. A panel of antibodies, CAM5.2, LH1, AE8, LP2K and LH8 recognising keratins 8, 10, 13, 19 and a basal cell marker respectively were employed. Keratins were identified using a standard immunocytochemical technique (Vectastain) and assessed on a 3 point scale, for both smears and biopsies. The vast majority of tumours were well differentiated. No particular keratin profile scen within the smear was associated with any particular state of differentiation. Although the sensitivity of K19 was greatest, its specificity was poor. The keratin antibodies with the best positive predictive value were CAM5.2 (K8) and the marker of the basal cell phenotype, LH8. The combination of down regulation of the secondary differentiation markers (K13, K10) coupled with 'simple' keratin expression (K8, K19) would seem to be the most consistent profile. We conclude that for exfoliative cytological screening to be of value as a diagnostic test it remains necessary to employ assays using more than one antikeratin antibody.

Topics: Biomarkers, Tumor; Biopsy; Carcinoma, Squamous Cell; Cytodiagnosis; Histocytochemistry; Humans; Keratins; Mouth Neoplasms; Predictive Value of Tests; Sensitivity and Specificity

Cytokeratins (CK), the intermediate filament markers for epithelial cells were analysed in 23 squamous cell carcinomas (SCC) of the tongue and 11 SCC of the alveolar mucosa (AM) by SDS-PAGE, immunoblotting and two dimensional gel electrophoresis. Normal human adult ventral tongue expresses CK nos 4, 5, 6, 13, 14, 16 (17) while the dorsal tongue expresses CK nos 1, 5, 6, 10, 14, 16 (17). CK 5 and CK 14 were not detected in a majority of samples and CK 18, a marker of simple epithelia, was aberrantly expressed in 18 samples. Normal human adult AM expresses CK nos 4, 5, 6, 13, 14, 16 (17). Among 11 SCC of AM, CK 4 and CK 5 were detected in only two samples each. CK 1 and CK 10 were aberrantly expressed in nine and one samples, respectively. The basic CKs such as CK 4, 5 and 14 were not expressed in SCC at both these sites while others like CK 1 and 18 were aberrantly expressed. Thus, non-expression of basic keratin, CK 5, of the oral lining epithelia and aberrant expression of simple epithelial keratins seem to be the major events in malignant transformation in the oral epithelia.

Topics: Adult; Alveolar Process; Carcinoma, Squamous Cell; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Humans; Immunoblotting; Keratins; Mouth Mucosa; Mouth Neoplasms; Neoplasm Proteins; Tongue Neoplasms

The effect of sodium lauryl sulphate (SLS) on cytokeratin (CK) gene expression in hamster cheek pouch epithelium was studied with a hybridohistochemical technique. Using specific human anti-sense RNA probes, the plausible hamster mRNA counterparts for these human CK mRNAs were localized by detection of heterologous hybrids. In comparison with normal epithelium, the expression and distribution pattern of CK mRNAs in the hamster cheek pouch were obviously changed after application of SLS. There was a decreased expression of CK mRNAs in the hyperplastic basal layer, and increased expression in the hypertrophic granular layer. Strikingly, hybridization with the human CK 18 cRNA probe revealed an additionally expressed CK mRNA in the SLS-treated epithelium that was not found in the untreated epithelium. The present study indicates that cRNA probes for human CK mRNAs can be used successfully, not only to distinguish between different hamster CK mRNAs but also to investigate changes in CK gene expression upon the induction of non-neoplastic and neoplastic alterations in the hamster cheek pouch model. This may help elucidate the molecular changes involved in epithelial pathologies.

Topics: Animals; Cheek; Cricetinae; Detergents; Epithelium; Gene Expression Regulation; Gene Expression Regulation, Neoplastic; Histocytochemistry; Humans; Hybridomas; Hyperplasia; Hypertrophy; In Situ Hybridization; Keratins; Male; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; RNA Probes; RNA, Antisense; RNA, Complementary; RNA, Messenger; Sodium Dodecyl Sulfate; Surface-Active Agents

In immunohistochemistry, it is well known that the majority of monoclonal antibodies to keratins work best on fresh frozen tissue specimens, yet in clinical practice most biopsies are routinely fixed in formaldehyde. This seriously limits the range of keratins that can be reliably assessed in retrospective studies (particularly where only rare archival material exists) and where subtle changes during tissue differentiation may be important. Antigen retrieval using exposure to microwave radiation is one technique that has been applied successfully to other tumour markers (e.g., p53). However, few papers have used this method when immunolabelling for keratins, in spite of the widespread use of antikeratin antibodies as markers of differentiation. The effect of keratin antigen retrieval using microwave processing was assessed on a range of oral mucosal biopsies, since the oral cavity displays a wide range of keratins. A panel of six well characterized antibodies was chosen: LP34 (Ck1, 5, 6, 18), LH1 (Ck10), LL025 (Ck16), A53 BA2 (Ck19), AE8 (Ck13), and E3 (Ck17). For each specimen, one piece was stored in liquid nitrogen and another piece fixed in formalin. Tissue sections were cut from each and, using the peroxidase avidin biotin technique, keratin expression was recorded for a frozen section, a dewaxed section, and a microwave-heated dewaxed section. Although overall there was a 25% improvement in identification of keratins after microwaving, some antibodies performed better than others. Given that keratins have been shown to be of value in tumour diagnosis, this study suggests that microwave processing of archival material can be a valuable adjunct to such analysis.

Topics: Antibodies; Biopsy; Humans; Keratins; Microwaves; Mouth Mucosa; Mouth Neoplasms

Epidermoid carcinomas were induced in hamster buccal pouches with use of 7.12 dimethylbenz[a]anthracene (DMBA). In five animals that served as tumor controls (Group 1), right buccal pouches were painted with DMBA (0.5% solution in mineral oil) thrice weekly for 14 weeks. In five animals (Group 2), right buccal pouches were painted with DMBA and reduced glutathione (GSH) was administered systemically by mouth. Five animals (Group 3) received vitamin E instead of glutathione. An additional 20 animals (Groups 4, 5, 6, and 7) were untreated, vehicle, glutathione, and vitamin E controls, respectively. Glutathione and vitamin E were given in doses of 10 mg/kg in 0.5 ml of mineral oil thrice weekly on days alternate to DMBA painting. Treatment by GSH and vitamin E reduced the number and size of tumors that were formed. Histopathologically, there were also fewer sites of dysplasia, carcinoma in situ, and early invasive epidermoid carcinoma than in the tumor control animals. The formalin-fixed and paraffin-embedded buccal pouch sections were stained immunohistochemically with use of monoclonal antibodies for cytokeratins. These included high-molecular-weight keratins (50,000-68,000 mol wt) 10, 13, and 8 (k10, k13, and k8, respectively). Oral carcinomas and dysplastic sites exhibited basal and suprabasal (spinous layer) high levels of k10, k13, and k8 staining. Treatment with GSH or vitamin E increased the suprabasal staining for high-molecular-weight keratins and reduced the protein expression for k10, k13, or k8. This pattern of staining was observed in dysplastic as well as in carcinoma sites. These results indicate that cytokeratin protein expression could contribute to a common biomarker analysis for chemoprevention.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Antidotes; Antioxidants; Carcinogens; Carcinoma in Situ; Carcinoma, Squamous Cell; Cheek; Cricetinae; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; Glutathione; Immunohistochemistry; Keratins; Male; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Vitamin E

We previously showed that keratin profiles can be of value in the diagnosis of oral cancer when using exfoliative cytology. In the future, they may form part of a screening program for oral cancer. This study evaluated the influence of long-term storage on keratin expression. Smears were collected from the clinically normal buccal mucosa and dorsal tongue of 22 patients. Half were stored in a refrigerator (5 degrees C) and half in a freezer (-70 degrees C). A total of 528 smears were collected. A panel of three antikeratin antibodies (LP34, AE8 and 1C7) was used to identify the preservation of keratin expression (graded as absent, few cells positive or many cells positive). The results for smears from dorsal tongue indicated that many cells were impermeable by the antikeratin antibodies. However, a satisfactory level of keratin immunoreactivity was observed in smears from buccal mucosa stored at -70 degrees C for over one year. Results for storage at 5 degrees C for both sites were inadequate after one month. Thus, smears from nonkeratinized oral sites may be stored at -70 degrees C for at least one year without a profound loss of keratin immunoreactivity, thus allowing examination of archival material.

Topics: Cold Temperature; Cytodiagnosis; Freezing; Humans; Immunohistochemistry; Keratins; Mouth Mucosa; Mouth Neoplasms; Specimen Handling; Time Factors

Retinoids are promising agents for therapy of squamous cancers. In vitro, retinoids decrease expression of differentiation markers in head and neck squamous carcinoma cells. Little information is available on effects of retinoids on head and neck squamous carcinoma cell xenograft growth in vivo. To address this issue, head and neck squamous carcinoma cells (line 1483) were established as xenografts in nude mice. Control tumors grew rapidly with doubling times of 4-6 days to mean volumes of 1696 mm3 after 24 days. Histological analyses indicated the formation of well-differentiated squamous carcinoma cells exhibiting pronounced stratification (basal and suprabasal cells) and keratinization (keratin pearls) with abundant stroma. Cytokeratin 19 expression was restricted to the basal cell layers, and cytokeratin 4 expression was abundant in suprabasal cells. Mice were treated daily with 30 mg/kg 9-cis retinoic acid, 20 mg/kg all-trans-retinoic acid, or 60 mg/kg 13-cis retinoic acid by p.o. gavage on a schedule of 5 days/week over 4 weeks. Low micromolar (1.48-3.67 microM) and nanomolar (200-490 nM) concentrations of 9-cis retinoic acid and all-trans-retinoic acid were measured in plasmas and xenografts, respectively, 30 min after dosing. Retinoid treatment produced a marked suppression of the squamous cell differentiation of tumor cells manifest by decreased keratinization, loss of stratification, and accumulation of basal cells. This was accompanied by large decreases in the number of CK4-positive cells and concomitant increases of CK19-positive cells. REtinoic acid receptor-beta expression was also increased by 2.9-9.7-fold after chronic retinoid treatment. 9-cis retinoic acid and all-trans-retinoic acid decreased tumor volumes by 23 +/- 5 (SE) and 19 +/- 3%, respectively (P < or = 0.05); 13-cis retinoic acid was inactive. These retinoids did not decrease the rate of exponential tumor growth but increased the latent period until exponential growth began. These studies demonstrate that retinoids do not universally decrease tumor growth but profoundly suppress squamous cell differentiation in vivo in this xenograft model.

Topics: Animals; Antineoplastic Agents; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Cell Division; Dose-Response Relationship, Drug; Female; Humans; Keratins; Mice; Mice, Nude; Mouth Neoplasms; Neoplasm Transplantation; Receptors, Retinoic Acid; Retinoids; RNA, Messenger; Transplantation, Heterologous; Tumor Cells, Cultured

The aim of the present study was to investigate epithelial cell proliferation in the linings of odontogenic cysts, including three different subtypes of odontogenic keratocyst (OKC), namely simple (non-recurrent), recurrent and basal cell naevus syndrome (BCNS)-associated lesions. Ki67 immunoreactivity in OKC (simple, n = 10; recurrent, n = 8; syndrome, n = 9), dentigerous cysts (DC, n = 5), radicular cysts (RC, n = 5) and normal oral mucosa (n = 7) was studied using a biotin-streptavidin-peroxidase method on paraffin sections after microwave treatment. Ki67+ epithelial cells were counted manually and related to the length of basement membrane (BM) as determined by TV image analysis. Data were analysed by the Mann-Whitney U test. The number of Ki67+ cells in simple OKC linings (53.1 +/- 17.8 cells/mmBM) was similar to that in oral epithelium (42.5 +/- 12.7 cells/mmBM; P > 0.2). However, both contained significantly more Ki67+ cells than DC (3.9 +/- 1.3 cells/mmBM) and RC linings (6.7 +/- 4.8 cells/mmBM; P < 0.006). The epithelial distribution of Ki67+ cells differed between groups, with the percentage of positive cells in basal layers in OKC linings (7.0 +/- 2.1%) being significantly lower than that of oral epithelia (35.9 +/- 5.6%), DC (78.4 +/- 8.4%) and RC (80.6 +/- 7.7%) linings respectively (P < 0.003). Comparison of Ki67 expression within the OKC group revealed no significant difference between simple and recurrent lesions (44.0 +/- 13.8 cells/mmBM; P > 0.3). However, OKC associated with BCNS contained significantly higher numbers of Ki67+ cells (91.8 +/- 35.6 cells/mmBM; P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Analysis of Variance; Basal Cell Nevus Syndrome; Biomarkers; Cell Cycle Proteins; Cell Division; Humans; Immunoenzyme Techniques; Keratins; Ki-67 Antigen; Microwaves; Mouth Neoplasms; Neoplasm Proteins; Nuclear Proteins; Odontogenic Cysts; Paraffin Embedding; Proliferating Cell Nuclear Antigen; Recurrence; S Phase; Statistics, Nonparametric

Serum-free cultures of normal human buccal epithelial cells were transfected with a plasmid containing the SV40 T-antigen (SV40T) gene. Two major lines developed that showed extended lifespans (between 30 and 40 weeks) as compared with the controls (approximately 6 weeks). Continued growth through one or two crises generated several sublines. They expressed the epithelial marker keratin and also exhibited nuclear expression of SV40T. The lines showed abnormal karyotypes with both numerical and structural aberrations and variably responded to agents that normally inhibit growth and/or induce terminal differentiation, i.e. transforming growth factor-beta 1 and fetal bovine serum. One of the lines, termed SVpgC2a, developed into an apparently immortal line, since it had undergone more than 700 population doublings from over 2 years in culture. Further characterization of this line demonstrated its clonal origin, with integration of two copies of SV40T at the same site and the presence of both normal retinoblastoma and wild-type p53 proteins. This line showed high resistance to growth inhibition by transforming growth factor-beta 1 and serum similar to that shown by buccal carcinoma cell line SqCC/Y1. Neither SVpgC2a nor its parental lines were tumorigenic when injected into athymic nude mice, whereas the SqCC/Y1 cells induced tumors. The various lines with extended but finite lifespans, complemented by one immortalized line, which retained non-malignant properties upon extended culture, provide a battery of model systems that will be useful for studying mechanisms of human oral carcinogenesis.

Topics: Animals; Antigens, Polyomavirus Transforming; Biomarkers; Blood; Cattle; Cell Differentiation; Cell Division; Cell Line; Cells, Cultured; Culture Media, Serum-Free; Epithelial Cells; Epithelium; Genes, Viral; Humans; Karyotyping; Keratins; Mice; Mice, Nude; Mouth Mucosa; Mouth Neoplasms; Recombinant Proteins; Retinoblastoma Protein; Simian virus 40; Transfection; Transforming Growth Factor beta; Transplantation, Heterologous; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Virus Integration

This article reports a series of 103 cases of giant cell fibromas occurring in the oral mucosa. The commonest location was the gingiva, followed by the tongue and the buccal mucosa. The mean age of the patients was 27.7 years, and the median age 21 years. Microscopically, the tumors were characterized by the presence of large stellate or angular cells, which occasionally contained several nuclei. Immunohistochemical stains showed that the cells were vimentin-positive but negative for S-100 protein, cytokeratin, leukocyte common antigen, and neurofilament.

Topics: Adolescent; Adult; Aged; Cell Nucleus; Child; Child, Preschool; Female; Fibroma; Giant Cells; Gingival Neoplasms; Humans; Immunohistochemistry; Keratins; Leukocyte Common Antigens; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neurofilament Proteins; S100 Proteins; Tongue Neoplasms; Vimentin

The well-documented specificity of anticytokeratin monoclonal antibodies for detection of epithelial micrometastatic cancer cells in bone marrow as a prognostic indicator inspired us to apply this approach to patients with squamous cell carcinomas (SSC) of the head and neck region. The sensitivity of the broad-spectrum anticytokeratin monoclonal antibody (mAb) A45-B/B3 used for tumor cell detection was demonstrated by immunostaining of cryostat sections from the respective primary tumors. Analysis of 31 patients with SSC revealed A45-B/B3-positive cells in 10 cases (32.3%) at frequencies of 1-207 per 1 x 10(6) mononuclear cells. Most specimens displayed isolated tumor cells, while cell clusters were found in only two cases (6.5%). The present data suggest that hematogenous dissemination of cancer cells is more frequent than expected from clinicopathologic staging of patients with SSC of the head and neck region.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Biomarkers, Tumor; Bone Marrow; Carcinoma, Squamous Cell; Coloring Agents; Epithelium; Female; Humans; Immunoglobulin G; Immunohistochemistry; Keratins; Lymph Node Excision; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Neoplastic Cells, Circulating; Prognosis; Sensitivity and Specificity

This study examined the expression of TGF-beta cell-surface receptors, the response to exogenous TGF-beta 1 and the autocrine production of TGF-beta in normal and squamous cell carcinoma-derived human oral keratinocytes with variable degrees of cellular differentiation. TGF-beta receptor expression, the response to exogenous ligand and the autocrine production of TGF-beta appeared unrelated to cellular differentiation. Cells expressed variable proportions of type-I, -II and -III TGF-beta receptors. The expression of type-III receptors correlated inversely with the expression of type-I receptors, but there was no relationship between type-II and either type-I or type-III TGF-beta receptors. Normal cells and the majority (7 of 8) of tumour-derived keratinocytes were inhibited by exogenous TGF-beta 1 and the degree of inhibition correlated with the expression of type-I, but not type-II or type-III, TGF-beta receptors. One tumour-derived cell line was refractory to exogenous TGF-beta 1 although it expressed all 3 receptor types. Endogenous TGF-beta was produced by both normal and tumour-derived keratinocytes and correlated inversely to the expression of type-I, but not type-II, TGF-beta receptors. Further, cells that produced more autocrine TGF-beta had a diminished response to exogenous TGF-beta 1. The data indicate a complex interaction between the expression of TGF-beta cell-surface receptors, endogenous ligand production and the cellular response to exogenous TGF-beta 1.

Topics: 3T3 Cells; Animals; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line; Culture Media, Conditioned; Humans; Keratinocytes; Keratins; Mice; Mouth Mucosa; Mouth Neoplasms; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta; Tumor Cells, Cultured; Vimentin

The effects of four different hyperplastic agents and of the carcinogen DMBA on cytokeratin expression in hamster cheek pouch epithelia were compared. Reversible hyperplasia was produced by the application of either oil of turpentine, vitamin A or TPA. No hyperplastic changes were produced by application of EPP. Apart from the transient appearance of a 45 kDa cytokeratin in one group treated with vitamin A, the immunohistochemical staining patterns and immunoblot profiles of cytokeratins from cheek pouches treated with each of the hyperplastic agents were identical to controls. Following application of DMBA, the cytokeratins stained with increased intensity in the spinous and granular cell layers. This was associated with increased amounts of 42-56 kDa cytokeratins and decreased production of 62-75 kDa cytokeratins. Monoclonal antibody AE1 detected a 45 kDa cytokeratin in extracts of DMBA-treated epithelia that was not detected in untreated epithelial extracts. Monoclonal antibody AE3 detected an additional 54 kDa cytokeratin band in extracts of DMBA-treated epithelia. These cytokeratin changes were present in preneoplastic epithelia and maintained in neoplastic epithelia.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Alkynes; Animals; Cricetinae; Diterpenes; Epithelium; Gene Expression Regulation, Neoplastic; Hyperplasia; Immunoblotting; Immunohistochemistry; Keratins; Male; Molecular Weight; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Retinyl Esters; Tetradecanoylphorbol Acetate; Turpentine; Vitamin A

Fifty 4- to 6-week-old male random-bred golden hamsters were injected intraperitoneally with a weight-related dose (12.5 mg/kg body weight) of N-methyl-N-nitrosourea (NMU) three times a week for 4 weeks. Groups of seven animals were killed 10, 16 and 22 weeks after the first injection. The palatal gingiva from six animals and the intermolar mucosa from 21 animals was examined. Seven male age-matched untreated control animals were killed at each period. Although all NMU-treated hamsters showed dysplastic and neoplastic changes similar to those in human oral squamous-cell carcinoma, other changes such as acantholytic dyskeratosis, invading cysts, duct-like structures and basaloid islands and cords were not. The extent and severity of the changes increased with time so that by 22 weeks there was extensive involvement of the palatal bone and marrow spaces, the molar periodontal ligament and the greater palatine neurovascular bundle by neoplastic epithelium. The invading epithelium was derived from the junctional, crevicular and palatal gingival and intermolar epithelium. The latent period for the crevicular and junctional epithelia was shorter than that for the palatal gingival and intermolar epithelium. The invasive changes from the latter epithelium were often preceded by exophytic changes such as epithelial projections, papillae and papillomas. Such changes were infrequent for the gingival, crevicular and junctional epithelia. The study shows that intraperitoneal NMU acts as a complete carcinogen on the palatal gingival and intermolar epithelium in hamsters.

Topics: Animals; Connective Tissue; Cricetinae; Epithelium; Gingiva; Gingival Neoplasms; Injections, Intraperitoneal; Keratins; Male; Mesocricetus; Methylnitrosourea; Molar; Mouth Mucosa; Mouth Neoplasms; Palate; Papilloma; Precancerous Conditions

The origin of the stromal, stellate and multinucleate cells in oral giant cell fibroma is unclear. Sixteen giant cell fibromas were stained immunocytochemically for keratin (MNF 116), vimentin, S-100 protein, neurofilaments, glial fibrillary acidic protein, alpha-smooth muscle actin, desmin, CD31 (PECAM-1), CD68, Factor XIIIa and prolyl 4-hydroxylase (5B5). In all cases positive staining was found with vimentin and prolyl 4-hydroxylase, indicating a functional fibroblast phenotype. Reactivity for Factor XIIIa was seen in two cases and in only one was a small number of giant cells stained, suggesting that the majority of oral giant cell fibromas are unrelated to the histologically similar fibrous papule of the nose or facial angiofibroma.

Topics: Actins; Adolescent; Adult; Aged; Animals; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Cell Adhesion Molecules; Desmin; Female; Fibroblasts; Fibroma; Giant Cell Tumors; Glial Fibrillary Acidic Protein; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Neurofilament Proteins; Phenotype; Platelet Endothelial Cell Adhesion Molecule-1; Procollagen-Proline Dioxygenase; S100 Proteins; Stromal Cells; Transglutaminases; Vimentin

Expression of cytoskeletal proteins has been shown to be dependent on the differentiation status of the tissue. In the present study, expression of two cytoskeletal proteins normally present in terminally differentiated keratinocytes, namely cytokeratin types 10/11 and involucrin, were studied in different stages of tumour progression in the oral mucosa. Results showed that cytokeratins 10/11 and involucrin strongly correlated with the differentiation status of cells. High expression was observed in non-dysplastic hyperplastic epithelium as compared to normal, dysplastic and neoplastic epithelium. In addition, various grades of dysplasia showed an inverse correlation with expression of these proteins. Statistical analysis of the results also showed a negative correlation between the differentiation of upper spinal cells and the stage of tumour progression. It therefore appears that the proteins studied may be useful as markers for epithelial carcinogenesis.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Cell Transformation, Neoplastic; Humans; Immunoenzyme Techniques; Keratinocytes; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Protein Precursors

Simple epithelial keratins K7, K8, and K18 are present in no more than trace amounts in normal stratified squamous epithelial but have been reported in squamous cell carcinomas. With the aim of determining the level at which keratin synthesis is regulated in vivo, we have compared the expression of mRNA by in situ hybridization and protein by immunohistochemistry for K7, K8, and K18 in a series of normal, dysplastic, and malignant oral epithelia. In normal epithelia mRNAs for K7, K8, and K18 were present in basal and lower spinous cells but adjacent sections were generally negative for the respective proteins. In severe dysplasia there was irregular suprabasal extension of K8 and K18 mRNAs in all cases and their proteins were expressed in more than half of the cases. The carcinomas expressed K8 and K18 mRNAs homogeneously and were strongly reactive for these keratin proteins but K7 expression appeared reduced in malignancy. These results are consistent with the post-transcriptional regulation of K7, K8, and K18 expression in normal epithelia and the presence of their proteins in dysplastic and malignant epithelia suggests the release of these epithelial cells from a post-transcriptional block on K8 and K18 translation. Alternatively, rapid degradation of K8 and K18 protein might be occurring in normal epithelia but be suppressed in dysplasia and malignancy.

Topics: Base Sequence; Carcinoma, Squamous Cell; Humans; In Situ Hybridization; Keratins; Molecular Sequence Data; Mouth Mucosa; Mouth Neoplasms; Oligonucleotide Probes; Reference Values; RNA, Messenger

The expression pattern of cytokeratin filaments in epithelia has been shown to be dependent on their type and grade of differentiation. The type of expression of cytokeratin in a cell may also be altered during carcinogenesis or other pathological conditions. The present study examined the alterations in expression of various cytokeratin types during different stages of tumour progression in the oral mucosa. Six monoclonal anti-cytokeratin antibodies were used for the study. Conspicuous staining differences with these antibodies were evident between normal keratinizing and non-keratinizing mucosa. These differences can be correlated to the differentiation pattern of the normal mucosa types. Antibodies specific to cytokeratin types 10/11, 19 and 14 showed significant correlation with stage of tumour progression. In addition cytokeratin type 18 also showed prominent differences in expression between different stages of tumour progression. These alterations in cytokeratin expression suggest that the terminal differentiation pathway of keratinocytes is disturbed during oral carcinogenesis. The results of the present study also emphasize the potential of using cytokeratin filaments as markers in the biological staging of oral premalignant and malignant lesions.

Topics: Antibodies, Monoclonal; Carcinoma; Carcinoma, Squamous Cell; Carcinoma, Verrucous; Humans; Immunohistochemistry; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms

Differences in gene transcription between RNA samples extracted from oral normal and squamous cell carcinoma (SCC) tissue were examined using the technique of cDNA library differential plaque screening. A differentially expressed transcript was selected on the basis of it being under-expressed in the cancer tissue and was identified, using DNA sequencing, as cytokeratin 14. The level of cytokeratin 14 transcription in RNA samples extracted from a range of oral SCC and normal tissue, as well as "white patch" lesions, was then investigated. Cytokeratin 14 appeared to be significantly under-expressed in oral cancer specimens studied compared to normal and white-patch tissue (P < 0.01). The trend for higher levels of cytokeratin 14 transcription in the dysplastic "white patch" samples compared to that observed for the malignant tissue (P < 0.05) suggests that the decrease in cytokeratin 14 transcription is a late event in the carcinogenic pathway.

Topics: Adolescent; Adult; Aged; Base Sequence; Carcinoma, Squamous Cell; DNA Probes; Female; Gene Expression; Gene Library; Humans; In Situ Hybridization; Keratins; Male; Middle Aged; Molecular Sequence Data; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; RNA, Messenger; Transcription, Genetic

Refinements in oral exfoliative cytology may make it a suitable screening technique for the early diagnosis of oral cancer. In this study DNA range profiles were combined with keratin expression in an attempt to improve the diagnostic accuracy of oral exfoliative cytology. Smears were taken from 33 biopsy-proven oral cancers and the contralateral normal site. For DNA range profiles the smears underwent Feulgen hydrolysis, with DNA distribution being assessed using the Vickers M85 microdensitometer. For keratin expression a panel of antikeratin antibodies were applied. The smears for keratin expression were then graded on a three-point scale. Abnormal DNA range profiles were observed in 23 of 33 smears taken from oral cancers and in two smears from normal oral mucosa (sensitivity 70%, specificity 90%, positive predictive value 90%). The simple epithelial keratins 8 and 19 were identified in the majority of oral cancer smears. The sensitivity of keratin 19 was greater (90%). However, keratin 8 was the most useful keratin marker associated with malignancy (sensitivity 62%, specificity 100%, positive predictive value 100%). The combination of simple keratin expression and DNA content improved the cancer detection rate beyond that obtainable with DNA range profile alone.

Topics: Adult; Aged; Aged, 80 and over; DNA, Neoplasm; Female; Humans; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Sensitivity and Specificity

In a retrospective pilot study we compared morphologically the first biopsies of 20 patients suffering from oral squamous cell carcinoma (OSCC) where preoperatively performed radiation therapy [gamma rays (cobalt-60)] was successful with 20 patients who underwent the same therapy without the expected success. All specimens were formalin fixed and paraffin embedded. We performed haematoxylin and eosin staining and immunohistochemistry (ABC-method) applying broad spectrum cytokeratin, cytokeratin (CK) 1 + 2, 3 + 6, 13 and anti-proliferating cell nuclear antigen (PCNA, PC10). The specimens of the patients with success of the radiotherapy showed statistically higher levels of PCNA positive tumour cells, measured by a computerised morphometric analysis system (VIDAS). These specimens showed significantly less CK 3-6 positive tumour cells than the specimens of the patients with failure of this therapy. The difference in the content of proliferating cell nuclear antigen might explain the different results of the performed radiation therapy. The difference in cytokeratin 3 + 6 expression might be linked with the different amount of benign hyperproliferation. Prospective studies are planned to prove the results.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Division; Female; Humans; Immunoenzyme Techniques; Keratins; Logistic Models; Male; Middle Aged; Mouth Neoplasms; Nuclear Proteins; Pilot Projects; Prognosis; Proliferating Cell Nuclear Antigen; Retrospective Studies; Treatment Outcome

Although cytokeratin expression is said to alter with the state of tumor differentiation, few studies appear to have confirmed this in fresh tissue biopsies from oral squamous cell carcinomas. Previous studies have been limited by the number of antibodies utilized, small sample size, or lack of information regarding tumor differentiation. A panel of 8 antikeratin antibodies (of which five recognised the simple epithelial keratins, 8, 18 and 19) were applied to fresh tissue biopsies from 24 oral cancer and 15 normal oral mucosal biopsies. A standard immunocytochemical technique was followed (Vectastain ABC method), with keratin expression graded on a 3 point scale. Our analysis indicated that simple epithelial keratins (K8, K18, K19) were not confined to the more poorly differentiated tumors. This may be relevant to tumor prognosis.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Keratins; Mouth Neoplasms; Neoplasm Proteins; Prognosis

A case of basaloid squamous carcinoma that involves the posterior buccal mucosa is described. The major histopathologic feature is a carcinoma with a basaloid pattern in association with squamous differentiation. The basaloid cells exhibit large and vesicular nuclei and eosinophilic clear or vacuolated cytoplasm. Cells are distributed in cords, trabeculae, or lobules that occasionally show glandular arrangement. The majority of the tumor cells are positive for keratin and a large group of cells distributed in glandular arrangement are positive also for vimentin.

Topics: Adult; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Humans; Keratins; Male; Mouth Mucosa; Mouth Neoplasms; Vimentin

To assess keratin profiles from smears of malignant and contralateral normal oral mucosa as part of the development of a screening procedure for oral cancer based on exfoliative cytology.. Smears were taken from oral cancers (confirmed by biopsy) and from the contralateral site of 20 patients. Using a panel of antikeratin antibodies, the keratins expressed by these cells were identified using a standard immunocytochemical technique (Vectastain) and assessed on a 3 point scale.. Using chi 2 analysis, noticeable differences between the keratin profiles for malignant mucosal smears compared with the contralateral mucosal smears were found. This was particularly evident for the simple epithelial keratins.. Individual keratins can be identified in smears from oral cancers. The identification of simple epithelial keratins seem to be the best keratin markers associated with malignancy. Their detection within smears from oral lesions could be valuable in the early diagnosis of oral cancer.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Humans; Immunoenzyme Techniques; Keratins; Mouth Mucosa; Mouth Neoplasms

The relationship between the histological grading of malignancy and the expression of vimentin and cytokeratin was studied in 43 cases of oral squamous cell carcinoma. Immunohistochemical analysis was carried out with the avidin-biotin method using monoclonal antibody anti-vimentin, and the peroxidase anti-peroxidase method using the polyclonal antibody anti-cytokeratin. All cases were classified according to the histological malignancy grading system proposed by Anneroth. All of the carcinomas were found to express cytokeratin, while 60.4% expressed vimentin. Vimentin was particularly noted in all tumors scored to have highly malignant cells.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Mouth Neoplasms; Prognosis; Vimentin

It was hypothesised that one may be able to visualise field changes, which are proposed to exist around tumours, as alterations in keratin intermediate filament protein expression. Standard immunohistochemical analysis using a panel of monoclonal anti-keratin antibodies was applied to fresh tissue sections to look for subtle changes in epithelial differentiation not visible in H&E sections. Such changes were observed in clinically normal epithelium from oral cancer patients, involving primarily substantial expression of keratins K8/K7 (using CAM 5.2) in the basal cells of 12 out of 34 biopsies, and also a trend towards a reduction in the complexity of keratin differentiation. Monitoring such changes may prove to be a valuable adjunct to conventional H&E staining if found to have prognostic and diagnostic significance.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cytoskeleton; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth; Mouth Neoplasms; Mucous Membrane

This study was undertaken to analyze keratin gene expression at both the mRNA and protein level in oral hairy leukoplakia (OHL). Comparisons were made with normal lingual epithelium from a similar site, tongue biting, normal buccal mucosa and another condition which disturbs oral epithelial differentiation, white sponge nevus. Combined immunocytochemical and in situ hybridization studies for keratins 14 and 19 were carried out on 2 specimens of OHL from HIV-positive males and one sample each of the other cases. Keratin 14 protein expression was uniform throughout all the epithelia. In normal epithelia and in lesions other than OHL, keratin 14 mRNA was most strongly expressed in basal cells with weaker but still significant amounts in the spinous cell layer. In both cases of OHL there was weaker basal cell expression of keratin 14 mRNA and frequent absence in koilocytoid cells. Keratin 19 protein expression was heterogeneous in the basal layer of all specimens with suprabasal staining of occasional groups of cells. Its mRNA was uniformly distributed in all cases. The findings indicate the keratin mRNA expression does not always parallel that of protein and that, in the case of keratin 14, expression may be influenced by the presence of EBV.

Topics: Adult; AIDS-Related Opportunistic Infections; Autoradiography; Basement Membrane; Epithelium; Female; Gene Expression Regulation; Herpesvirus 4, Human; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Leukoplakia, Oral; Male; Middle Aged; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Nevus; RNA, Messenger; Tongue Diseases; Tongue Habits; Tumor Virus Infections

The expression of cytokeratins (CK) 1, 4, 5/6, 8, 13, 18, 19 and 20 and involucrin in 42 cases of squamous cell carcinomas from various locations was examined. The tumours expressed CK5/6 in 55%, CK8 in 76%, CK13 in 43% and CK19 in 95% of cases. The CK5/6-positive primary tumours were from uterine cervix, head and neck, lung, skin, oesophagus and urinary bladder, and the CK13-positive primary tumours were from uterine cervix, lung and vulva. Metastatic squamous cell carcinomas from head and neck more frequently expressed CK5/6 and 13, 7/7 (100%) and 6/7 (86%) compared with 3/5 (60%) and 0/5 (0%) in the primary squamous cell carcinomas. Few cases were CK1, CK4 and CK18 immunoreactive. CK20 immunoreactivity was not observed. Involucrin was expressed in 71% of tumours, and most of the involucrin-positive cells were located at the central parts of tumour cell clusters except for one case in which the peripheral cells around tumour cell clusters were positive. Thus, expression of the so-called simple epithelial markers CK8 and CK19 occurs in the majority of squamous cell carcinomas. The absence of CK20 immunoreactivity may be helpful in differential diagnosis.

Topics: Carcinoma, Squamous Cell; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Mouth Neoplasms; Protein Precursors; Skin Neoplasms; Urinary Bladder Neoplasms; Uterine Neoplasms

Only five cases of basaloid squamous cell carcinoma (BSCC), a rare tumor of head and neck, have been reported to involve the floor of mouth.. Clinicopathologic and immunohistochemical features of eight BSCC of floor of mouth were studied to evaluate the significance of the basaloid features.. Five patients were male and three were female. Their mean age was 52 years (range, 39-59). At presentation, one patient was diagnosed with Stage II disease, four were diagnosed with Stage III disease, and three were diagnosed with Stage IV disease. Aside from typical squamous differentiation, each patient had a component of basaloid cells arranged in irregular nests, cords, or pseudoglandular spaces with a brisk mitotic rate, myxoid stroma, and marked tendency for perineural invasion. A panel of immunostains yielded the following results: keratin, +8/8; carcinoembryonic antigen, +3/8; and S-100, chromogranin, and neuron-specific enolase were negative. Mucin stains were negative in all cases. Ultrastructural characterization of three BSCC revealed squamous differentiation of the basaloid cells and a peculiar basal membrane-like material in between them. No neurosecretory granules were present. Seven patients underwent surgery; six of them were also treated with postoperative radiation therapy. In two cases, chemotherapy was added at recurrence. One nonresectable patient received radiation and chemotherapy. At the last follow-up, five patients were dead of disease within 13 months from the diagnosis. One patient died of an unknown cause. Two patients were still alive at the time of this report, 4 and 2 months after treatment. Seven patients had recurrent disease. The authors compared these data with a control group of patients with conventional squamous cell carcinoma (SCC).. The authors' results indicate that BSCC of floor of mouth is an aggressive variant of SCC and is prognostically worse than the conventional SCC, regardless of the grade of the latter.

Topics: Adult; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Female; Humans; Keratins; Male; Middle Aged; Mouth Floor; Mouth Neoplasms; Neoplasm Staging; Prognosis; Survival Rate

A spontaneous squamous cell carcinoma was diagnosed in the oral cavity of an adult female squirrel monkey (Saimiri sciureus). Immunohistochemical analysis of the neoplasm demonstrated cytokeratin and vimentin, but not S100 or desmin in the neoplastic epithelial cells.

Topics: Animals; Carcinoma, Squamous Cell; Desmin; Female; Immunohistochemistry; Keratins; Monkey Diseases; Mouth Neoplasms; S100 Proteins; Saimiri; Vimentin

The expression of differentiation associated high PM Keratin polypeptides of the oral mucosa lesions were studied by immunohistochemical and immunoblotting techniques applied to adjacent sections of each biopsy specimen. The material studied included specimens of leukoplakia, verrucous carcinoma, squamous cell carcinoma, adenocarcinoma and keratoacanthoma. Little or no expression of 65-67 Kd keratins was evident in squamous cell carcinoma and adenocarcinoma. Hyperkeratotic (both benign and dysplastic) lesions such as verrucous carcinoma, leukoplakia, and keratoacanthoma, showed great variations in the intensity of 65-67 bands and a very irregular immunohistochemical staining pattern. Increased amounts of horny substance was usually accompanied by absence of, or decreased expression of 65-67 Kd keratins, thus indicating a change in the polypeptide composition of the horny layer in pathological conditions of the oral epithelium.

Topics: Adenocarcinoma; Antigens, Differentiation; Carcinoma, Squamous Cell; Carcinoma, Verrucous; Cell Differentiation; Electrophoresis, Polyacrylamide Gel; Humans; Immunoblotting; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Keratoacanthoma; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms

An expression of desmosomal glycoprotein 1 (DG 1) was immunohistochemically examined in 77 biopsies and 21 metastatic cervical lymph nodes of oral squamous cell carcinomas (SCC). In the primary tumors the DG 1 expression was significantly reduced at the invasive site of poorly differentiated and highly invasive tumors. In cases of metastases in cervical lymph nodes, the DG 1 staining at the invasive site of the primary tumor was significantly less than that of nonmetastatic cases. The DG 1 expression in the metastatic lymph nodes was as weak as that in the primary tumor. Thus, we suggest that immunohistochemical investigation of DG 1 expression in oral SCC is valuable in predicting tumor behavior.

Topics: Carcinoma, Squamous Cell; Cell Adhesion Molecules; Cell Transformation, Neoplastic; Cytoskeletal Proteins; Desmoglein 1; Desmoplakins; Desmosomes; Humans; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Lymphatic Metastasis; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Staging; Staining and Labeling

The various cytokeratin polypeptides in oral epithelia are expressed in dependence on site and formation of a stratum corneum. Certain cytokeratins occur permanently and others occasionally. In fibrous hyperplasia and Lichen ruber planus, patterns of cytokeratins did not deviate significantly from normal. In some but not all cases of squamous cell carcinoma and leukoplakia studied, marked aberrations of pattern were characterized by (i) appearance of cytokeratin No. 19, (ii) somewhat more frequent occurrence of cytokeratins Nos. 8 and 18, (iii) proteolytic modifications of cytokeratins, and (iv) partial loss of a few site-specific cytokeratins. The aberrations may be taken as additional diagnostic criteria for differentiation between non-aggressive and potentially aggressive leukoplakic lesion, even if they are not correlated with the conventional histological grading of dysplasia.

Topics: Adolescent; Adult; Aged; Carcinoma, Squamous Cell; Female; Gingiva; Humans; Hyperplasia; Keratins; Keratosis; Leukoplakia, Oral; Lichen Planus; Male; Middle Aged; Molecular Weight; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms

Cellular expression of keratins (intermediate filaments) can be demonstrated by immunocytochemistry using unfixed tissue. However, for practical reasons, provision of fresh tissue to the laboratory is often difficult. Recently a fresh tissue transport medium (Patent applied for) has been developed which allows keratin immunocytochemistry to be performed up to 4 days after biopsy. In this study oral mucosal biopsies from 10 patients were hemisected, half placed in the new transport medium at 4 degrees C and the other half immediately frozen in liquid nitrogen. After 4 days frozen sections of both halves of the biopsies were stained by immunocytochemistry using various antikeratin antibodies. The morphology and staining characteristics of the two halves of the biopsies were then assessed. No significant difference could be found in either morphology or preservation of keratin expression in specimens stored in transport medium, as compared to those in liquid nitrogen. This new transport medium may offer considerable advantage for the provision of a histologic and immunocytochemical diagnostic service.

Topics: Cacodylic Acid; Carcinoma, Squamous Cell; Culture Media; Freezing; Gelatin; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratins; Leukoplakia, Oral; Lichen Planus; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Nitrogen; Specimen Handling; Staining and Labeling

The keratin expression pattern in oral stratified epithelium is related to the cellular differentiation level. The normal pattern shows the keratin pair K5 and K14 in the stratum basale whereas K1 and K10, or K4 and K13, are the two pairs associated with differentiating suprabasal cells. Monoclonal and polyclonal antibodies to individual keratins (K10, K13 and K14) were used in a two-color immunofluorescence staining method to study their coexpression in single cells. Altered keratin expression in premalignant and malignant lesions indicated abnormal differentiation. Monospecific keratin antibodies were suggested to be useful for evaluation of epithelial differentiation changes in oral dysplasias and oral squamous cell carcinomas.

Topics: Biomarkers, Tumor; Carcinoma in Situ; Carcinoma, Squamous Cell; Epithelium; Fluorescent Antibody Technique; Humans; Keratins; Leukoplakia, Oral; Microscopy, Fluorescence; Mouth Mucosa; Mouth Neoplasms; Photography; Precancerous Conditions

An 8-year-old girl with a non-familial case of white sponge nevus (WSN) is presented. The differential diagnosis is discussed with reference to anamnestic, clinical and histopathological data. In keratin expression, WSN resembles the epithelia of the hard palate and the tongue.

Topics: Child; Female; Humans; Immunoenzyme Techniques; Keratins; Mouth Mucosa; Mouth Neoplasms; Nevus; Telangiectasis

A survey of 4342 oral pathology reports accumulated over a five-year period was performed. Diagnoses were 1090 irritation fibromas and 116 giant cell fibromas. A statistical comparison was then made between the giant cell fibromas and the irritation fibromas to determine if there were any differences between these two lesions with respect to sex or race predilection, age distribution, or location in the oral cavity. Finally, various staining techniques were performed on the giant cell fibromas in an attempt to ascertain the origin of the giant cells present in these lesions. The results will be discussed in this paper.

Topics: Adolescent; Adult; Aged; Child; Child, Preschool; Collagen; Epithelium; Female; Fibroma; Giant Cells; Gingival Neoplasms; Humans; Hyperplasia; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Mouth Neoplasms; Palatal Neoplasms; Tongue Neoplasms

Recent studies have suggested that the epithelial expression of two leukocyte-related proteins, human class II HLA-DR antigen and myelomonocytic L1 antigen, depends on a certain state of cellular maturation and differentiation. We have studied HLA-DR and L1 expression in oral squamous cell carcinomas. The epithelial distribution of these proteins was evaluated in relation to differentiation alterations by two-color immunofluorescence staining with cytokeratins (K14 and K13) as a baseline. HLA-DR was infrequently expressed in oral carcinomas, apparently being unrelated to the degree of differentiation and the subepithelial leukocyte infiltration. L1 was generally present in oral epithelium but disappeared in the most invasive cells of carcinomas. These cells were also K14 and K13 negative suggesting an abnormal state of differentiation. L1 has been suggested to have an inhibitory effect on casein kinases I and II, enzymes possibly associated with cell proliferation; it might therefore exert an inhibitory effect on tumor growth. Its absence could be an interesting aspect of the invasiveness of oral carcinoma cells.

Topics: Antigens, Surface; Carcinoma, Squamous Cell; Cell Adhesion Molecules, Neuronal; Cell Differentiation; Epithelium; Fluorescent Antibody Technique; HLA-DR Antigens; Humans; Keratins; Leukocyte L1 Antigen Complex; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness

Recognition of premalignant lesions in the oral epithelium has the potential to increase survival rates for squamous cell carcinoma of the oral cavity. It has previously been reported that cytokeratin 19 (CK19), a 40-kd epithelial cytoskeletal protein within the suprabasal squamous epithelium, is a specific marker of moderate-to-severe dysplasia and carcinoma in situ in oral cavity squamous epithelium. In contrast, normal epithelium and hyperplastic lesions reportedly express CK19 only in the basal layer if at all. The authors chose to test and extend this hypothesis by studying suprabasal CK19 expression and dysplasia of the oral cavity and upper aerodigestive tract in paraffin-embedded specimens that had been fixed in alcohol, a superior fixative for the preservation of cytokeratins. The authors examined 56 alcohol-fixed, paraffin-embedded specimens including 37 from the oral cavity, using two antibodies specific for CK19 (Ks19.1 and 4.62), an antibody to the nuclear proliferation marker, proliferating cell nuclear antigen (PCNA) (19A2), and an antibody to the putative tumor suppressor gene, p53 (pAb1801). The lesions were classified as normal, hyperplasia, mild dysplasia, moderate dysplasia, severe dysplasia/carcinoma in situ, or invasive squamous cell carcinoma, following standard histologic criteria. Immunocytochemically stained sections were scored for the presence or absence of suprabasal CK19, suprabasal PCNA, and p53 positivity, regardless of location. The immunostaining patterns of the two anti-CK19 antibodies were essentially equivalent. Except for one laryngeal specimen, normal epithelium, when positive, showed CK19 expression only in scattered cells throughout the basal layer. Proliferating cell nuclear antigen-positive nuclei were found exclusively in the basal layer. In areas of hyperplasia, CK19 immunostaining was absent or confined to the basal layer in 20 of 38 specimens and was expressed in suprabasal cells in 18 of 38 hyperplastic specimens. Proliferating cell nuclear antigen immunostaining in all cases of hyperplasia was limited to the basal layer. Severe dysplasia and carcinoma in situ showed suprabasal CK19 staining in six of nine specimens and no CK19 staining in three of nine specimens. In contrast, suprabasal PCNA immunostaining was found in all dysplasia and carcinoma in situ cases. p53 expression was detected in three of nine severe dysplasia/CIS specimens and was immunocytochemically undetectable in all normal, hyperplasia, an

Topics: Antigens, Neoplasm; Carcinoma, Squamous Cell; Genes, p53; Humans; Hyperplasia; Immunohistochemistry; Keratins; Mouth; Mouth Mucosa; Mouth Neoplasms; Mutation; Nuclear Proteins; Pharynx; Precancerous Conditions; Proliferating Cell Nuclear Antigen; Tissue Fixation

We have analyzed the expression of the three retinoic acid receptor (RAR) (alpha, beta, gamma) mRNAs and the intermediate filament protein keratin 19 (K19) mRNA in cell lines cultured from oral and epidermal human squamous cell carcinoma (SCC) and from benign, hyperplastic, and hyperkeratotic (leukoplakia) lesions arising in various regions of the oral cavity. Seven of the SCC lines were derived from tumors arising in regions of the oral cavity in which the normal epithelial cells (keratinocytes) express RAR beta transcripts. Seven of the nine SCC lines tested did not exhibit detectable RAR beta mRNA levels, even in response to addition of retinoic acid (RA). The RAR beta gene did not appear to be rearranged or deleted in the five nonexpressing SCC lines examined by Southern analysis. The steady-state RAR gamma mRNA levels were 2- to 4-fold lower in 6 of the 9 SCC lines than in their normal counterparts, whereas the RAR alpha message levels in SCC lines were similar to those of the normal cell strains. The expression of keratin 19 message, which is RA inducible in normal keratinocytes, was also abnormal in many of the SCC cell lines. Some SCC lines, e.g., those derived form tumors of the soft palate epithelium, did not express high levels of K19 message even though normal soft palate keratinocytes expressed high levels of K19 mRNA. Two of the nine SCC lines expressed higher than normal levels of K19 mRNA, and this expression was RA independent. Cells cultured from four oral leukoplakia lesions were also examined and found to express RAR beta mRNA at relatively normal levels, but they expressed RAR gamma message at half the level of epithelial cells cultured from normal tissue. These results show that the correlation between RAR beta gene expression and K19 gene expression that we have observed in the various normal keratinocyte subtypes of the oral cavity (D.L. Crowe et al., manuscript in preparation) is not present in transformed keratinocytes (SCC cells). The lack of apparent RA regulation of the K19 gene in SCC lines may be associated with other aberrations in differentiation which have been identified in SCC cells. Abnormally low expression of the RAR beta receptor may contribute to neoplastic progression in stratified squamous epithelia. It may also determine whether a tumor is responsive to RA as a chemotherapeutic agent.

Topics: Blotting, Southern; Carcinoma, Squamous Cell; Carrier Proteins; DNA; Genomic Library; Humans; Keratinocytes; Keratins; Leukoplakia; Mouth Neoplasms; Receptors, Retinoic Acid; RNA, Messenger; Skin Neoplasms; Transcription, Genetic; Tumor Cells, Cultured

Four cases of adenosquamous carcinoma from the oral and pharyngeal cavities were analyzed by light microscopy and immunohistochemistry. Lymph node metastases were present in three cases. One patient died 2 years after treatment. All four carcinomas presented a mixture of squamous and glandular mucus-secreting neoplastic elements. Immunostaining for high-molecular-weight cytokeratins (KL1) was constantly positive in both squamous and glandular tumor cells. Antibodies against low-molecular-weight cytokeratins (K19) and carcinoembryonic antigen were positive only in the glandular component. The histological aspect and the immunohistochemical phenotype of these tumors is similar to the ordinary squamous cell carcinoma and adenocarcinoma, respectively.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Humans; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Pharyngeal Neoplasms

Six cases of spindle cell squamous carcinoma (SCSC) of the oral cavity were studied clinicopathologically, immunohistochemically and ultrastructurally to summarize the clinicopathological features of this rare neoplasm and to discuss the debatable histogenesis of the sarcomatoid component and the differential diagnosis of SCSC. The mean age of the patients was 72 years and the female to male ratio was 1:2. Four of them had a history of irradiation for pre-existing squamous cell carcinoma. One patient died of SCSC. While clinical and histological prognostic factors of SCSC could not be determined, it was shown that radical surgery resulted in good prognosis. The epithelial nature of the sarcomatoid component of SCSC was clearly revealed by a combination of immunohistochemical staining for keratins and electron microscopic demonstration of tonofilament-like filaments and/or desmosome-like structures. Together with electron microscopic evaluation of the tumour cells, immunohistochemical characterization of tumour cells using antibodies to keratin, vimentin, glial fibrillary acidic protein and S-100 protein is very helpful in differentiating SCSC from true spindle cell sarcoma, melanoma and malignant myoepithelioma. In the immunohistochemical differential diagnosis of SCSC, it is important to remember that SCSC should not be ruled out of the differential diagnosis by a positive reaction for vimentin in sarcomatoid tumour cells. Absence of staining for keratin in the sarcomatoid tumour cells does not always exclude SCSC, because some SCSCs show immunoreactivity of keratin in their sarcomatoid components only with some anti-keratin antibodies. Different kinds of anti-keratin antibodies should be applied in the differential diagnosis of SCSC.

Topics: Aged; Aged, 80 and over; Carcinoma; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Male; Microscopy, Electron; Middle Aged; Mouth Neoplasms; Vimentin

Precancerous lesions and squamous cell carcinomas (SCCs) were induced in the oral mucosa of outbred male Sprague-Dawley rats by repeated application of the carcinogen 4-nitroquinoline-1-oxide. Temporal alterations in the pattern of cytokeratins expressed by epithelial cells in these developing neoplasms were investigated by means of one- and two-dimensional polyacrylamide gel electrophoresis and by probing electrophoretic blots of these gels with anticytokeratin antibodies. Progressive diminution in the expression of a 58.3 kDa cytokeratin was detected in moderately dysplastic epithelium and subsequent lesions, as was reduced expression of a 53.5 kDa cytokeratin after a stage of severe dysplasia had been induced. Analysis of two-dimensional electrophoretograms indicated an alkaline shift of acidic variants of a 49.0 kDa cytokeratin in moderately dysplastic epithelium; this shift was more pronounced in the induced, severely dysplastic epithelium and SCCs. The latter finding of an alkaline shift in cytokeratins of dysplastic epithelial cells has not been previously reported in preneoplastic lesions.

Topics: 4-Nitroquinoline-1-oxide; Animals; Biomarkers, Tumor; Carcinoma, Squamous Cell; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Keratins; Male; Mouth Mucosa; Mouth Neoplasms; Rats; Rats, Inbred Strains; Time Factors

Supplementary prognostic factors should be added to the TNM classification for oral squamous cell carcinomas in order to optimize its clinical value. We have recently published two prognostically valuable malignancy grading systems based on histopathology and immunohistology of the most invasive cells in oral squamous cell carcinomas (OSCCs). However, a major problem with classifications based on histologic features is frequent lack of interobserver agreement which limits the clinical value of subjective histologic classifications. Thirty-eight file cases of OSCCs were therefore graded by three pathologists according to criteria of the histologic malignancy grading system which includes 5 morphologic features, each graded from 1 to 4. Agreement was calculated by kappa statistics, which showed that interobserver agreement was not optimal, but significantly better than by chance alone. We also studied the reproducibility of grading of immunohistologic membrane expression of a tumor-associated marker (blood group antigen H), and found a similar level of agreement. We conclude that the clinical value of our grading systems will increase by improving reproducibility.

Topics: Carcinoma, Squamous Cell; Cell Membrane; Cell Nucleus; Humans; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Lymphocytes; Mitosis; Mouth Neoplasms; Multivariate Analysis; Neoplasm Invasiveness; Observer Variation; Plasma Cells; Polymorphism, Genetic; Prognosis; Reproducibility of Results; Survival Rate

Four cell lines were established from squamous cell carcinomas (SCC) of the oral cavity. Cell lines AW 13516 and AW 8507 were derived from poorly differentiated SCC and epidermoid carcinoma of the tongue respectively. Cell line AW 10498 was derived from moderately differentiated SCC of the lower alveolus, and AW 9803 grew from a well-differentiated SCC of a retromolar trigone. The cultures showed typical epithelial cell morphology, numerous mitotic figures, occasional multinucleated giant cells, individual cell diskeratosis and nuclear and nucleolar abnormalities. The cell lines AW 13516 and AW 8507 were fast growers with a doubling time of 35.5 h and 31.9 h, respectively, which was independent of the initial seeding density. Cell lines AW 10498 (doubling time 52.2 h) and AW 9803 (doubling time 66 h) showed slower growth and had shorter doubling times at higher seeding densities. The presence of cytokeratins was detected in all the four cell lines by using polyclonal antikeratin antisera in indirect immunofluorescence and in Western blotting. None of the cell lines expressed major histocompatibility complex (MHC) class II antigens. MHC class I antigens were expressed by three cell lines but not by AW 9803. Flow cytometric analysis of DNA content and chromosomal studies suggested the presence of polyploidy and aneuploidy in all the four cell lines. Ultrastructural studies revealed typical epithelial cell features, such as the presence of desmosomes, tonofilaments and keratin bundles.

Topics: Carcinoma, Squamous Cell; DNA, Neoplasm; Flow Cytometry; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Humans; Keratins; Mouth Neoplasms; Tumor Cells, Cultured

Two cell lines from head-and-neck squamous-cell carcinomas (SCC) have been established and characterized. Cell line R105 was derived from a xenografted SCC of the floor of the mouth and cell line T87/rc from a SCC of the epiglottis. Identification of individual cytokeratins 4, 5, 7, 8, 10, 13, 14, 18 and 19 led to the conclusion that both cell types had squamous characteristics and that keratinization occurred in xenografts. Ultrastructurally, junctional complexes were observed in both cell lines. Characteristic marker chromosomes were found and although both cell lines were derived from male patients, the Y chromosome was missing from all examined cells. The basic biological parameters of both cell lines were modal chromosome numbers of 59 (R105) and 60 (T87/rc), a doubling time of 60 (R105) and 45 hr (T87/rc) and a DNA index of 1.54 (R105) and 1.31 (T87/rc). The tumorigenicity of the 2 cell lines was proved by the ability to form colonies on a plastic substratum, as well as in a soft agar assay. Furthermore, the cells could produce multi-cellular tumour spheroids, and formed tumour nodules after subcutaneous inoculation into nude mice. The R105 tumour cells appeared to be better differentiated than the T87/rc as observed by histology and immuno(histo)chemistry. Both cell lines appear to retain SCC differentiation after being xenografted into nude mice, cultured for more than 40 passages in vitro and thereafter again xenografted into nude mice.

Topics: Animals; Antibodies, Monoclonal; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line; Desmin; Fluorescent Antibody Technique; Humans; Karyotyping; Keratins; Mice; Microscopy, Electron; Mouth Neoplasms; Transplantation, Heterologous; Vimentin

This study examined the initial behaviour of 48 human oral squamous cell carcinomas (SCC) in cell culture. The early outcome of these cultures (contamination, absence of cell growth, epithelial cell senescence/fibroblast overgrowth, extended keratinocyte growth) did not reflect the clinical characteristics of the tumours of origin. Four new human oral SCC cell lines were characterized more extensively. Each cell line was immortal, 3T3-independent, and expressed low degrees of anchorage independence (CFE less than 4 per cent). Two of the four cell lines were tumorigenic in athymic mice. All of the cell lines expressed keratin intermediate filaments and two showed weak co-expression of vimentin. A wide range of keratins were expressed by the tumour xenografts; cornified keratins (K1, K10) were only expressed in the absence of K19 and vimentin, and vice versa. The nuclear:cytoplasmic ratio and the degree of serum independence correlated with each other and with the STNMP clinical grading of the tumours of origin.

Topics: Adult; Aged; Aged, 80 and over; Animals; Carcinoma, Squamous Cell; Cell Count; Cell Line; Female; Fibroblasts; Humans; Keratins; Male; Mice; Mice, Nude; Middle Aged; Mouth Neoplasms; Neoplasm Transplantation; Tumor Cells, Cultured; Vimentin

This study was undertaken to explore the expression of keratins in the hamster cheek pouch carcinogenesis model, using monospecific keratin antibodies and a technique that allows immunoblotting analysis of tissues embedded in paraffin. Changes in keratin expression were correlated with histopathological changes and with the expression of the enzyme gamma-glutamyl transpeptidase. The right cheek pouch of 20 male golden Syrian hamsters was treated with 0.5% 7,12-dimethylbenz[a]anthracene for 16 weeks. As previously described by other laboratories, this treatment resulted in hyperplastic and dysplastic lesions and benign and malignant tumors. The keratins assayed in this study were K14 (Mr 55,000), K1 (Mr 67,000), and K13 (Mr 47,000). The normal hamster cheek pouch epithelium expressed K14 in the basal layer and K13 in the suprabasal and differentiated layers, whereas K1 was not detected by either immunohistochemistry or immunoblotting. Concomitant with 7,12-dimethylbenz[a]anthracene-induced hyperplasia, there were some topographical alterations in the distribution of K14. In this case, K14 was no longer restricted to the basal layer but was also expressed in differentiated cells. The same pattern was also observed in dysplastic lesions and in squamous cell carcinoma. Furthermore, expression of the K13 differentiation-associated keratin was preserved in this hyperplastic epithelium during all the stages of carcinogenesis, including either anaplastic or differentiated areas. In contrast, after 2 weeks of 7,12-dimethylbenz[a]anthracene treatment, K1 expression started as a weak and patchy pattern in suprabasal cells, becoming stronger and more homogeneous at 8 and 16 weeks of treatment. However, K1 was almost absent in squamous cell carcinoma, where only small very well differentiated areas were stained. We also observed gamma-glutamyl transpeptidase-positive foci in earlier stages of carcinogenesis, concomitant with the expression of the K1 keratin. However, it was not possible to find a perfect topographical correspondence between the two events. Alterations in the pattern of keratin expression appear to be a common feature during the development of squamous cell carcinoma in different systems and could be an excellent tool to study carcinogenesis and chemoprevention.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Biomarkers, Tumor; Carcinogenicity Tests; Carcinoma, Squamous Cell; Cell Differentiation; Cheek; Cricetinae; Hyperplasia; Keratins; Male; Mesocricetus; Molecular Weight; Mouth Mucosa; Mouth Neoplasms; Reference Values

The well and poorly differentiated oral squamous carcinomas preferentially express proteins, blood group antigens, and contain associated dendritic Langerhans' cells. Keratin pearls in well-differentiated carcinomas simulate the differentiation pathway of the normal oral squamous epithelium, whereas poorly differentiated carcinomas do not and appear more heterogeneous. Terminally keratinized cells correlate with involucrin and expression of blood group antigens in keratin pearls, a feature that differs from the nonkeratinizing normal epithelium in which such carcinomas arise. Dendritic Langerhans' cells are reduced in number in squamous carcinomas.

Topics: ABO Blood-Group System; beta 2-Microglobulin; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Dendritic Cells; Fibronectins; HLA Antigens; Humans; Keratins; Laminin; Langerhans Cells; Mouth Neoplasms; Protein Precursors; S100 Proteins

Topics: Adult; Cell Differentiation; Epithelium; Female; Fluorescent Antibody Technique; Humans; Intestinal Mucosa; Jejunum; Keratins; Male; Microsurgery; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Postoperative Complications

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Drug Evaluation; Female; Humans; Immunoenzyme Techniques; Isotretinoin; Keratins; Male; Micronucleus Tests; Middle Aged; Mouth Neoplasms; Protein Precursors; Transglutaminases

Recently, regional changes of cytokeratin patterns in human normal non-keratinized or keratinized oral mucosa have been demonstrated and the expression of individual cytokeratin polypeptides in lesions of oral mucosa has been compared with that of normal tissues. In particular, the presence of cytokeratin 19 in the suprabasal cell layers of oral epithelia has been shown to be strongly correlated with premalignancy. In the present study, we describe the results of an immunohistochemical investigation performed using a monoclonal antibody specific for cytokeratin 1 on normal oral mucosa and benign or malignant oral lesions. We show the different distribution of this polypeptide in non-neoplastic lesions from different sites of oral mucosa and describe the presence of cytokeratin 19. Our results are in agreement with the data obtained previously. In the malignant cases we demonstrate that the distribution of the two cytokeratins is characterized by complementary patterns.

Topics: Carcinoma, Squamous Cell; Histocytochemistry; Humans; Immunoblotting; Immunoenzyme Techniques; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Papilloma; Precancerous Conditions; Tissue Distribution

The hamster cheek pouch (HCP) serves as an excellent model system not only for the studies on initiation and promotion but also for the modulation of experimental oral carcinogenesis. In our studies, HCPs treated with 7,12-dimethylbenz[a]anthracene (DMBA) showed both cheek pouch and stomach papillomas. Utilizing this model system, we tested and compared the modulatory effects of snuff, retinoic acid, and beta-carotene on the incidence of tumors and the keratin expression pattern. HCPs treated with snuff, either alone or in combination with DMBA, resulted in stomach papillomas. HCPs treated with snuff showed no cheek pouch tumors, and those treated with snuff and DMBA showed only 10-15% tumor incidence. Both beta-carotene and retinoic acid showed a total inhibition of DMBA-induced carcinogenesis in the HCP as well as in the stomach. The keratin expression pattern showed alterations depending on the experimental conditions.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; beta Carotene; Carotenoids; Cheek; Cricetinae; Keratins; Male; Mesocricetus; Mouth Neoplasms; Plants, Toxic; Tobacco, Smokeless; Tretinoin

High-grade carcinoma of the oral cavity is considered to be an aggressive malignancy associated with a poor prognosis. The tumors are often considered to be relatively radiosensitive. Because previous reviews of oral cavity cancer have contained few patients with grade 4 lesions, little is known about the true behavior of these tumors. Information was collected on 80 patients with high-grade carcinoma of the oral cavity to determine the clinical findings and optimal treatment of this tumor. Primary tumor location, presence of neck metastasis, and clinical stage did not affect survival. Tumor size was a significant risk factor. The 5-year survival rate for patients with high-grade carcinoma treated by combination surgery and radiation (52%) was significantly greater (p less than or equal to 0.05) than that for patients treated by radiation alone (28%) or surgery alone (24%). This study is the first to describe the clinical course of a large number of patients exclusively with high-grade carcinoma of the oral cavity.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Combined Modality Therapy; Epidermal Cells; Female; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Metastasis

This study examined the effect of 3T3 fibroblasts on the expression of anchorage independence and the degree of cornification in early cultures of three carcinoma-derived epithelial cell lines (R59, R63a, R63b) and in one cell line derived from non-malignant dysplastic epithelium where there was no evidence of invasion (R66a). The epithelial cell lines originated from the palatal (R63a, R66a) and the lingual (R59, R63b) mucosa of rats that had been painted with the carcinogen 4-nitroquinoline N-oxide. In the absence of 3T3 fibroblasts, progressive culture resulted in an increase in the colony forming efficiency (CFE) of R63a, R63b and R59 and a decrease in the percentage of cornified cells in all cell lines. 3T3 fibroblasts caused a decrease in the CFE and the degree of cornification in the 3T3-dependent cell line (R63a), particularly at the lower passages, but these parameters remained essentially unchanged by 3T3 fibroblasts in the 3T3-independent cell lines (R59, R63b). 3T3 fibroblasts did not influence the cornification of R66a and this cell line remained anchorage dependent throughout the study. The results suggest that in malignant cell lines characterised by being independent of 3T3 fibroblasts (R63b, R59) the CFE was inversely correlated to the degree of cornification. However, in the malignant cell line showing a greater dependence on support (R63a) the relationship between CFE and cornification was unclear because these parameters may have been modulated by the presence of 3T3 fibroblasts. The cell line from dysplastic non-invasive tissue (R66a) differed from its malignant counterparts in the fact that CFE and cornification were unaffected by 3T3 fibroblasts despite previous studies showing a dependence on mesenchymal support.

Topics: Animals; Cell Adhesion; Cell Line; Epidermal Cells; Epithelial Cells; Fibroblasts; Keratins; Male; Mouth Mucosa; Mouth Neoplasms; Rats; Rats, Inbred Strains

Epithelial dendritic cells (EDC) were examined in human oral tissues with non-specific keratosis, lichen planus and squamous cell carcinoma. Acetone-fixed frozen sections were stained using an indirect immunoperoxidase technique and monoclonal antibodies to the human CD1 thymocyte (OKT6) and HLA-DR antigens. Significantly more T6+ and DR+ EDC were present in lichen planus tissues than normal controls, tissues with non-specific keratosis and the epithelial overlying/adjacent to squamous cell carcinomas, the latter tissues having comparable numbers of both T6+ and DR+ EDC. By contrast, significantly fewer T6+ EDC and significantly more DR+ cells were present in the invasive epithelium of squamous cell carcinomas than the overlying/adjacent epithelium of carcinomas, the non-specific keratosis group and the normal tissues. 23-60% of pathological tissues had either focal or general DR+ reactivity in keratinocytes, but there was no correlation between the density of T6+ or DR+ EDC and the keratinocyte DR status of the tissues. The results suggest that immunological enhancement occurs in lichen planus and possibly immunological impairment may characterize invasive squamous cell carcinoma.

Topics: Carcinoma, Squamous Cell; Dendritic Cells; Epidermis; Epithelium; HLA-DR Antigens; Humans; Immunoenzyme Techniques; Keratins; Langerhans Cells; Leukoplakia, Oral; Lichen Planus; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms

Five cases of monophasic and 7 cases of biphasic spindle-cell carcinomas were analyzed immunohistochemically for the presence of vimentin and keratin type intermediate filaments in the pleomorphic spindle cells. Vimentin reactivity proved to be a consistent feature but keratin reactivity was more variable, this latter filament being lost in two cases initially presenting as pure squamous cell carcinomas showing dedifferentiation towards a pure monophasic spindle-cell tumour when recurring. The converse was also noted: acquisition of keratin in a monophasic spindle-cell tumour that recurred as squamous cell carcinoma. These results were considered to support the concept that spindle-cell tumours of the upper aerodigestive tract are a peculiar type of carcinoma and not a product of a pluripotent stem cell exhibiting bidirectional differentiation. Diagnostic implications are as follows: keratin positivity in a spindle-cell tumour substantiates its carcinomatous nature but its absence does not rule out a diagnosis of spindle-cell carcinoma.

Topics: Aged; Aged, 80 and over; Carcinoma; Carcinoma, Squamous Cell; Female; Humans; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Neoplasm Recurrence, Local; Vimentin

Cytokeratin pattern was analyzed in 14 moderately differentiated and 12 well-differentiated squamous cell carcinomas of buccal mucosa by SDS-PAGE, immunoblotting and two dimensional electrophoresis. These were compared with patterns of normal buccal mucosa and surrounding areas whenever possible. Normal buccal mucosa expresses keratin No. 4 (59Kd), 5 (58Kd), 13 (54Kd) and 14 (50Kd). Keratin No. 4 (59Kd) and 14 (50Kd) were expressed by 20 of 26 tumors studied, while many of the tumors did not express keratins No. 5 (58Kd) and 13 (54Kd). Keratin No. 1 (67Kd) and 16 (48Kd) were aberrantly expressed by 9 well-differentiated tumors. Keratin No. 17 (46Kd) and 18 (45Kd) were expressed by 10 and 8 tumors of 14 moderately differentiated tumors. Six tumors which showed involvement of alveolar mucosa, expressed some keratins expressed by its normal counterpart. Their altered expression was consistent with the differentiation pattern as stated earlier. Non-expression of keratins 5 and 13 seems to be the result of malignant transformation and is seen in the majority of tumors, while appearance of aberrant keratins seems to be related more to the degree of differentiation of the tumor.

Topics: Carcinoma, Squamous Cell; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Humans; Immunoblotting; Isoelectric Focusing; Keratins; Mouth Mucosa; Mouth Neoplasms; Sodium Dodecyl Sulfate

The pattern of keratin expression in hamster cheek pouch epithelium during 15-wk of DMBA-induced carcinogenesis was studied. The sequential changes in cytokeratins of premalignant and malignant tissues and comparative investigation of normal epithelial tissues were examined during a weekly sequential DMBA-induced chemical carcinogenesis. Keratin polypeptides of normal pouch epithelium appear in a molecular weight range of 43-67 kd and 5-6 proteins can be identified. The disappearance of high molecular weight keratin (61-67 kd) was observed from the 6-wk DMBA-treated premalignant group to the 15-wk DMBA-treated malignant group. An additional keratin polypeptide was noted initially on the 11th-wk-DMBA-treated group and remained to the 15th-wk-DMBA treated group.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Cheek; Cricetinae; Electrophoresis, Polyacrylamide Gel; Epithelium; Immunoblotting; Keratins; Male; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Palate; Precancerous Conditions; Skin; Sodium Dodecyl Sulfate

The frequency of the occurrence of cytokeratins analyzed SDS-electrophoretically in 20 leukoplakic lesions and 14 squamous cell carcinomas of the oral mucosa has shown a picture of "restlessness" with some quantitative, some qualitative deviations from the locally normal pattern of cytokeratins. In most cases the basic pattern typical for the oral mucosa was still recognizable, except for three highly undifferentiated carcinomas. The variations of the cytokeratin pattern existed independently of the clinical form of leukoplakia and of its histological degree of dysplasia. The striking findings in some, but not all cases were: --the presence of cytokeratin no. 18 as a major component which normally appears rarely and faintly, and of cytokeratin no. 19, which is not normally detectable electrophoretically within the whole oral mucosa; --the absence of cytokeratins no. 1-3 despite of the cornification which was reliably proved by histology; --the appearance of the proteins having molecular mass values of 42 kDa and less which very probably may be the products of partial keratinolysis as evidenced by immunoblotting with monoclonal and polyclonal antibodies to cytokeratins. Among these proteins a proteolytically modified cytokeratin no. 19 of only 38 kDa was found.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Electrophoresis, Polyacrylamide Gel; Humans; Immunoblotting; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Sodium Dodecyl Sulfate

Transforming growth factors have a wide range of biological activities related to cell proliferation and differentiation. In general TGF-alpha promotes cell proliferation while TGF-beta may stimulate or inhibit proliferation depending on the cell type and growth factor environment. Cultured human keratinocytes, skin and oral squamous cell carcinomas were analysed for the presence of transcripts and protein for the transforming growth factors alpha & beta. Both growth factors were detected in cultured keratinocytes (which have receptors for and respond to both ligands), and in medium conditioned by these cells. Additionally transcripts for TGF-alpha were found preferentially in the basal, proliferative compartment of cultured keratinocytes. Similarly both growth factors were detected in oral squamous cell carcinomas and a highly significant inverse correlation was found between the levels of TGF-alpha and the epidermal growth factor receptor in these tumours. The data for TGF-alpha are consistent with the existence of an autocrine growth control loop influencing cell proliferation in both a normal cell type and malignant epithelial tissues, a process that in keratinocytes and responsive squamous cell carcinomas could be modulated by TGF-beta.

Topics: Carcinoma, Squamous Cell; Cell Division; Cells, Cultured; Epidermal Cells; Epidermal Growth Factor; Epithelial Cells; Humans; Keratins; Mouth Neoplasms; Protein Precursors; Skin; Transforming Growth Factors; Tumor Cells, Cultured

As the distribution pattern of cytokeratin (CK), filaggrin and involucrin has recently been suggested to discriminate between benign and malignant epithelial growths, biopsies of healthy oral mucosa, leukoplakias without and with dysplasia and squamous cell carcinomas were examined immunohistochemically using a panel of 4 monoclonal antibodies (AB) against different cytokeratin polypeptides (34 beta E12, KL1 and Pkk1) and filaggrin as well as a polyclonal AB to involucrin. Major and statistically significant differences were observed in the profiles of CKs (except Pkk1), filaggrin and involucrin between leukoplakias without and with epithelial dysplasia. However, the alteration in the expression of CKs, filaggrin and involucrin proved to be not a constant feature in leukoplakias with dysplasia as a considerable portion (20-25%) of them revealed the profiles of CKs, filaggrin and involucrin similar to those of benign leukoplakias, and vice versa. Immunostaining of these antigens did not define the diagnosis of dysplasia in leukoplakias more precisely than grading in conventional histology can do so far. However, immunohistochemical sensitivity in detecting a broad range of variation in the abnormal maturation patterns of keratinocytes in leukoplakias with dysplasia can be used to divide these lesions into subgroups to elucidate their prognosis in follow-up studies.

Topics: Carcinoma, Squamous Cell; Epithelium; Female; Filaggrin Proteins; Humans; Hyperplasia; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratins; Leukoplakia, Oral; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Phosphoproteins; Protein Precursors

The evolution of squamous epithelial neoplasms induced by 7,12-dimethylbenz[a]anthracene (DMBA) in Syrian hamster buccal pouch epithelium (HBPE) and the angiogenic potential of a subpopulation of presumptive preneoplastic keratinocytes was evaluated by examining the ability of whole cell dissociates of HBPE and subpopulations of keratinocytes, or their 72-h serum-free conditioned media (CM), to induce neovascularization in rat corneas and directional migration of bovine adrenal gland capillary endothelial cells (BCE) in culture. Buccal pouches were treated in vivo twice weekly for 5 weeks with either DMBA, paraffin oil (PO) or received no treatment. Hamsters were killed at various times after the last application of carcinogen and single-cell suspensions were prepared by enzymatic dissociation. Angiogenesis was assayed by injecting HBPE cells, or by implanting Hydron pellets containing CM in corneas and observing directional ingrowth of capillary blood vessels. Directional migration of BCE under agarose was tested with CM. Angiogenic activity of DMBA-initiated HBPE dissociates was detected initially at 4 and 5 weeks after treatment, was markedly depressed between weeks 8 and 16 and re-emerged in squamous papillomas at week 25. The pattern of expression of angiogenic activity was observed to parallel the frequency of development of a morphologically unique population of keratinocytes that was detected exclusively in cultures of DMBA-exposed HBPE. These unique cells, designated type II keratinocytes, potently stimulated neovascularization in vivo and directional migration of BCE in culture. These results demonstrate that angiogenic activity is an early manifestation of hamster pouch carcinogenesis and suggests that type II keratinocytes, presumptive preneoplastic cells in this model, are the principal source of this activity.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Cell Separation; Cells, Cultured; Cheek; Cricetinae; Epithelium; Keratins; Mesocricetus; Microscopy, Phase-Contrast; Mouth Neoplasms; Neovascularization, Pathologic; Phenotype; Precancerous Conditions

The cheek pouch of the Syrian hamster is an excellent tissue for the experimental induction of oral cancer by carcinogenic chemicals. Lysate prepared from a cell line (HCPC-1) derived from one of these hamster oral tumors greatly increased the growth of these oral tumor cells in vitro. We now show that the mitogenic substance, transforming growth factor alpha (TGF-alpha), is present in all of the chemically transformed hamster oral tumors examined (in vitro and in vivo). In no adult normal tissue of the Syrian hamster can we detect expression of TGF-alpha. TGF-alpha could be partly or wholly responsible for the mitogenic activity detected in the lysate of the chemically transformed hamster oral keratinocytes. Both normal and chemically transformed hamster oral keratinocytes express the receptor to epidermal growth factor. The consistent detection of TGF-alpha and epidermal growth factor receptor mRNAs in these hamster oral tumor cells suggests that an autocrine growth mechanism might be operative. This hamster cheek pouch oral cancer model can be used for the molecular analysis of how TGF-alpha and epidermal growth factor receptor might be involved in the malignant transformation of epithelial tissues.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Squamous Cell; Cell Line; Cell Transformation, Neoplastic; Cloning, Molecular; Cricetinae; DNA Replication; Epidermal Cells; Epidermal Growth Factor; Epidermis; ErbB Receptors; Growth Substances; Keratins; Mouth Neoplasms; Peptides; Transforming Growth Factors

The hamster cheek pouch is an excellent target tissue for the experimental study of oral carcinogenesis. In the course of searching for molecular alterations during the malignant transformation process, the necessity for a molecular marker for cellular proliferation became apparent. In this report, we show that the cellular level of the histone H3 mRNA is valid as a molecular index of proliferation for cycling cell populations. H3 is known to be proliferation dependent for its expression in cultured animal cells. This study shows that H3 retains its cell-cycle-dependent expression in chemically transformed oral keratinocytes. The onset of H3 mRNA synthesis couples to the onset of DNA synthesis (S-phase). The cellular level of H3 mRNA therefore is proportional to the fraction of cells in the S-phase of the cell cycle. This conveniently allows us to correlate, in asynchronized cell populations, the expression of cellular genes to their proliferation rates. We demonstrate the usefulness of this proliferation marker by presenting data that different chemically induced oral carcinomas, but not normal cheek pouch tissues, contain readily detectable levels of c-Ki-ras proto-oncogene mRNA. Probing the same RNA blot to quantitate H3 mRNA levels allowed us to conclude that the high levels of c-Ki-ras mRNA in tumor tissues was likely due to the increased growth rate of the tumor tissues and not due to the deregulated expression of this cellular-proto-oncogene.

Topics: Animals; Blotting, Northern; Carcinoma, Squamous Cell; Cell Cycle; Cell Division; Cell Transformation, Neoplastic; Cricetinae; Epidermis; Gene Expression Regulation; Histones; Interphase; Keratins; Mouth Neoplasms; Proto-Oncogenes; RNA, Messenger; RNA, Neoplasm; Tumor Cells, Cultured

Normal rat oral keratinocytes have been transformed with the carcinogen 4-nitroquinoline N-oxide (4NQO) in vitro. The morphology, growth characteristics, ability to grow without anchorage and tumorigenicity in athymic mice was examined in 12 selected cell lines. Each of the lines could be assigned to one of two general groups. The first group of cell lines although showing some morphological signs of transformation and the ability to be subcultured beyond passage 15 were not anchorage independent or able to form tumours in athymic mice. The second group of cell lines showed distinct signs of morphological transformation, could be serially subcultured without 3T3 feeder cells, were anchorage independent and tumorigenic in athymic mice. Anchorage independence was more common at higher passages and with increased 4NQO treatment and correlated well with a decreased reliance on 3T3 feeder cell support. The anchorage-independent phenotype was closely associated with the ability to form tumours in athymic mice. This same sequence of phenotypic changes has been demonstrated in rat oral keratinocytes after 4NQO treatment in vivo indicating that during carcinogenesis, cell populations progress through the same stages whether proliferation occurs in vitro or in vivo. There is some evidence to suggest, however, that the time interval between stages may be altered when carcinogenesis takes place in vitro.

Topics: 4-Nitroquinoline-1-oxide; Animals; Cell Transformation, Neoplastic; Cells, Cultured; Epidermal Cells; Epidermis; Keratins; Male; Mouth Mucosa; Mouth Neoplasms; Nitroquinolines; Rats; Rats, Inbred Strains

Two cell lines designated HBPC-1 and HBPC-2 have been established from hamster buccal pouch tumors induced by topical 7,12-dimethylbenz(a)anthracene (DMBA) and DMBA in conjunction with type 1 herpes simplex virus infection, respectively. The cells are epithelial in morphology, have a doubling time of approximately 18 h, and require bovine serum for optimal growth. The karyotype is aneuploid, with several marker chromosomes, and the cells produce squamous cell carcinomas when transplanted into normal hamster pouch tissues.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Squamous Cell; Cell Division; Cheek; Cricetinae; Desmosomes; Epithelium; Keratins; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Neoplasm Transplantation; Simplexvirus; Tumor Cells, Cultured

Topics: Antibodies; Antibodies, Monoclonal; Biomarkers; Humans; Hyperplasia; Immunohistochemistry; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions

The altered pattern in the expression of keratin proteins as a function of tumour progression was studied in the hamster cheek pouch and compared the changes with electron microscopic observations using DMBA as a carcinogen. In the case of hyperplasia and well developed tumours a conspicuous loss of 67 K and an increase in 46 K was observed compared to the controls. The increased expression of low molecular weight keratins during tumour growth is well supported by the enhanced expression of tonofilament bundles, electron microscopically. This study suggests a triangular relationship between the presence of low molecular weight keratins-enhanced expression of tonofilament bundles and the undifferentiated nature of the oral tumours.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Cheek; Cricetinae; Cytoskeleton; Disease Models, Animal; Gene Expression Regulation; Intermediate Filaments; Keratins; Male; Mesocricetus; Microscopy, Electron; Mouth Neoplasms

Individual keratin proteins were identified in 10 DMBA induced squamous cell carcinomas (SCC) of the hamster cheek pouch. SDS-polyacrylamide gel electrophoresis of water-insoluble cytoskeletal extracts from the tumor tissue demonstrated alterations in the protein distribution normal for the site. Immunoblot analysis with a broad spectrum polyclonal antikeratin antiserum identified the keratins in the preparations and confirmed changes in their distribution in the tumor preparations. The major keratin species for all the tumor tissues ranged in molecular weight from 45 to 57kd. The normal tissues had keratins with molecular weights from 45 to 73kd. The absence of high molecular weight keratins was a prominent feature in all the cancers. The histologic appearance of the tumors was varied but the distribution of the keratins was not correlated with the various histologies. The results demonstrate that changes in keratin gene expression occur in DMBA-induced cheek-pouch carcinomas but the precise alterations in the keratin proteins from those seen normally are not predictable.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Squamous Cell; Cheek; Cricetinae; Gene Expression Regulation; Keratins; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Neoplasm Proteins

Two mouse monoclonal antibodies (MoAbs), B10 and 1H5, were generated by fusing mouse myeloma NS-1 cells with spleen cells from a BALB/c mouse immunized with Ueda-1 cells derived from human squamous cell carcinoma (SCC) of the floor of the mouth. Immunohistochemical analysis revealed that these MoAbs recognize the filamentous components of cytoplasm which were protein in nature. While the pattern of antigen distribution in various cell lines was not cell-type specific, reactivity of these antibodies with tissue sections was informative. MoAb 1H5 was preferentially reactive with well-differentiated squamous cell carcinoma, however, reaction with adenocarcinoma was observed infrequently. This antibody also preferentially reacted with the spinous layer of normal stratified squamous epithelium. MoAb B10, however, was reactive with nonepithelial tissues as well as with epithelial ones, and its level of binding bore no relationship to the grade of histologic malignancy. SDS-PAGE and Western blotting analysis, using cytokeratin extracts of Ueda-1 cells and human epidermis, demonstrated that MoAb B10 reacted with a wide range of keratin proteins of 65-67K, 58K, 56.5K, 56K, 52K, 50K, 48K, 45K, 40K, 38K, 36K, and 34K molecular weight (MW), while MoAb 1H5 reacted with keratin proteins of 65-67K, 58K, 56.5K, 56K, 52K, 48K, and 34K MW. These results suggest that MoAb 1H5 may recognize keratin subfamilies related to squamous differentiation, whereas MoAb B10 recognizes a wide range of keratin proteins, and may even react with other kinds of intermediate filament proteins (IFPs).

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Cell Line; Humans; Immunoenzyme Techniques; Keratins; Mouth Neoplasms

A total of 211 cases of benign and malignant tumors of epithelial origin were studied by the immunoperoxidase method to determine the distribution profile of epithelial membrane antigen (EMA) with the use of monoclonal antibody. Normal epithelial cells in the oral mucosa and skin were usually negative for EMA staining, as were epithelial cells in hyperplastic lesions and papillomas. Paget cells and tumor cells of Bowen's disease (carcinoma in situ) demonstrated a high incidence of EMA positivity, whereas the frequency in basal cell carcinoma was unexpectedly low. Squamous cell carcinomas revealed positive EMA staining of cytoplasmic membranes, and the antigen was also present in keratinized areas. EMA expression in squamous cell carcinoma generally showed a high incidence (85%) and was higher in keratinized lesions than in unkeratinized or less well-differentiated neoplasms. EMA distribution could be classified into two forms: one in which the cytoplasmic membranes demonstrate positivity and in which a positive cytoplasmic pattern is found in parakeratinized cells in malignant foci.

Topics: Antibodies, Monoclonal; Antigens; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Membrane Glycoproteins; Mouth Mucosa; Mouth Neoplasms; Mucin-1; Protein Precursors; Skin Neoplasms

Different tumours of the head and neck were analysed by immunohistochemistry. The distribution pattern of several intermediate filaments was studied. Keratin filaments were typical of carcinomas, whereas vimentin filaments were typical of mesenchymal tumours of different origin. The advances of this new technique of "tumour typing" are discussed.

Topics: Carcinoma, Basal Cell; Cytoskeleton; Diagnosis, Differential; Head and Neck Neoplasms; Humans; Hypopharyngeal Neoplasms; Immunoenzyme Techniques; Intermediate Filaments; Keratins; Lymphoma; Melanoma; Mouth Neoplasms; Nasopharyngeal Neoplasms; Neuroma, Acoustic; Oropharyngeal Neoplasms; Tonsillar Neoplasms; Vimentin

Involucrin has been recognized recently as a marker of terminal differentiation of squamous epithelial cells and also as a useful marker for keratinization; its expression in epithelial tumors of oral and pharyngeal mucosa and skin was examined. Involucrin in normal oral mucosa and skin was restricted to the granular and upper spinous layers and was absent in the basal layer. Hyperkeratosis was characterized by strong positive staining for involucrum in spinous and granular cell layers. A similar pattern was noted in seborrheic keratosis and verruca vulgaris. Condyloma acuminatum specimens revealed slight staining, whereas Paget cells were negative. Calcifying epitheliomas of Malherbe were usually unreactive. Papillomas exhibited the regular distribution of involucrin, as found in normal squamous epithelium. Basal cell carcinomas were generally negative, whereas squamous cell carcinomas showed an irregular distribution of involucrin. Immunohistochemical staining for involucrin may be useful for identification of keratinizing cells in epithelial tumor foci, just as is the use of monoclonal antibody to keratin KL1.

Topics: Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Papilloma; Pharyngeal Neoplasms; Protein Precursors; Skin Diseases; Skin Neoplasms; Staining and Labeling

Papillomas (40) and squamous cell carcinomas (75) were examined for the presence of three keratin proteins with the use of an immunohistochemical technique. Polyclonal keratin antibody (TK, detecting 41 to 65 kDa keratin) and monoclonal antibodies KL1 and PKK1 (55 to 57 kDa and 41 to 56 kDa, respectively) were used. Squamous epithelium in normal oral mucosa showed marked TK staining in cells of upper strata and relatively slight staining in basal layer cells, moderate KL1 staining in spinous and granular layers and was negative in basal cells. Positive PKK1 staining was noted in cells of the basal layer. Columnar epithelium in the nasal mucosa showed TK staining in all layers, KL1 staining on the apical side of epithelial cells and trace or negative staining in basal layer cells. There was moderate PKK1 staining along the apical side of cells and variable staining in basal cells. Keratin distribution in oral papillomas was similar to that in normal oral epithelium, whereas in nasal and nasopharyngeal papillomas, keratin distribution was restricted to the upper layers. Tonsillar papillomas showed a strong TK reaction, negative KL1 in upper layer cells, and marked PKK1 staining in basal cells. Well-keratinized squamous carcinomas indicated an irregular TK distribution and decreased KL1 and negative PKK1 stainings. Intermediate and poorly differentiated keratinizing squamous carcinoma showed irregular staining patterns for the three classes of keratins studied. Immunohistochemically detectable keratins utilizing monoclonal antibodies were described as useful markers of epithelial tumors of squamous origin. Keratin expression within benign tumors was related to normal regional distribution, whereas in malignant tumors, keratin distribution was irregular in its distribution profile.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Esophageal Neoplasms; Humans; Keratins; Mouth Mucosa; Mouth Neoplasms; Nasal Mucosa; Nasopharyngeal Neoplasms; Papilloma

An in vivo model of oral epithelial carcinogenesis in rats has been established successfully in cell culture. Oral carcinomas of the tongue and palate were induced in Sprague-Dawley male rats by painting their palates three times weekly for 7-8 months with 0.5% (w/v) 4-nitroquinoline-N-oxide. Oral keratinocytes from malignant and untreated control tissues were cultivated using 3T3 fibroblast support. Both the normal and malignant cells stained positively with an anti-human keratin polyclonal antibody but malignant keratinocytes were heterogeneous with regard to cell size, shape and intercellular packing, unlike the regular organization of the normal cultures. Malignant keratinocyte cultures differed markedly from their normal counterparts by an increase in their growth rate, the capacity for serial cultivation to the 25th passage (to date) and independence of 3T3 fibroblast support. In contrast, cultures established from healthy tissue showed signs of senescence usually by passage 4 and were totally reliant on 3T3 fibroblast support for growth. Malignant keratinocytes expressed anchorage independence when cultured in a semi-solid medium and gave rise to tumour formation in athymic mice. The development of this specialized cell culture system substantially increases the potential of the rat 4NQO model to investigate the pathogenesis of oral squamous cell carcinomas.

Topics: Animals; Carcinoma, Squamous Cell; Cells, Cultured; Epidermis; Keratins; Male; Mice; Mice, Nude; Mouth Neoplasms; Neoplasm Transplantation; Rats; Rats, Inbred Strains

A neoplastic epithelial cell line, TYS, was isolated from a well-differentiated squamous cell carcinoma expressing carcinoembryonic antigen (CEA) that arose in human oral mucosa. Expressions of CEA and amylase as well as ample tonofilaments were detected in cultured TYS cells. Transplantation of the cells into athymic nude mice resulted in production of adenoid squamous cell carcinoma containing CEA and amylase. Cultivation of TYS cells in the presence of sodium butyrate resulted in suppression of cell growth and production of secretory granules with amylase in the cytoplasm of the cells. When the sodium butyrate-treated cells were transplanted into nude mice, a small mass developed transiently at the inoculation site and then disappeared. This mass was histopathologically interpreted as acinic cell carcinoma with squamoid lesion. These findings suggest that we have established a human adenoid squamous carcinoma cell line presumably derived from a minor salivary gland present in oral mucosa.

Topics: Adenoids; Aged; Aged, 80 and over; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Cell Line; Female; Humans; Keratins; Lymphatic Diseases; Mouth Neoplasms; Neoplasm Transplantation; Neoplasms

Topics: Animals; Antibodies; Diagnosis, Differential; Face; Facial Neoplasms; Fluorescent Antibody Technique; Histological Techniques; Humans; Immune Sera; Keratins; Microscopy, Fluorescence; Mouth; Mouth Neoplasms; Rabbits

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Squamous Cell; Cricetinae; Humans; Keratins; Male; Mouth Mucosa; Mouth Neoplasms

Histological grading of squamous cell carcinoma is subjective and suffers from poor observer reproducibility. We investigated the feasibility of quantifying histological differentiation via point counting, using both the degree of keratinization and a novel definition of differentiation that was based on architectural features of the tumour. Multiple recounts of 20 cases of human oral squamous cell carcinoma were performed at several magnifications (X100, X160 and X250). Six lines of human squamous cell carcinoma tumour lines were examined for changes in differentiation following transplantation to athymic nude mice. Observer reproducibility was extremely high for all recounts except at the highest magnification, where the tumour architecture may have been obscured. Of the human squamous cell carcinomas transplanted to nude mice, five of six tumour lines showed significant histological changes, most commonly toward decreased differentiation. The changes were usually present in the initial transplant and were similar to those we have reported for transplants of adenocarcinomas. We conclude that histological differentiation can be quantified in squamous cell carcinomas with a high degree of observer reproducibility, even in the absence of keratinization; the method employed is sufficiently sensitive to be applied to practical problems of biological significance.

Topics: Animals; Carcinoma, Squamous Cell; Humans; Keratins; Mice; Mice, Nude; Mouth Neoplasms; Necrosis; Neoplasm Transplantation

Topics: Humans; Hyperplasia; Immunoenzyme Techniques; Keratins; Mouth Mucosa; Mouth Neoplasms; Pharyngeal Neoplasms

Topics: Carcinoma, Squamous Cell; Clinical Enzyme Tests; Diagnosis, Differential; Histocytochemistry; Humans; Keratins; Lymphoma; Mouth Neoplasms

Carcinosarcomas are rare tumors of the upper aerodigestive tract, consisting of both carcinomatous and sarcomatous tissue. The larynx and oral cavity are most frequently involved. There has been much controversy regarding the histological nature and clinical course of these tumors. We report a case of carcinosarcoma of the floor of mouth in a 76 year old man who presented with a large pedunculated sublingual mass. There was no evidence of regional or systemic metastatic disease. After local excision, he was followed for one year without evidence of recurrence or metastasis. A review of the literature is presented, with an attempt to clarify clinically relevant aspects of nomenclature, pathogenesis, and clinical course.

Topics: Aged; Carcinoma, Squamous Cell; Carcinosarcoma; Cell Transformation, Neoplastic; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Male; Microscopy, Electron; Mouth Floor; Mouth Mucosa; Mouth Neoplasms; Neoplasms, Multiple Primary

In oral dysplasias and squamous cell carcinomas, relationships exist between the presence of keratin filaments and cell differentiation. The keratinized areas of high differentiated carcinomas are carcinoembryonic antigen (CEA) positive. The labeling of the higher molecular keratins is similar to the distribution of lectin receptors so that lectins represent membrane-oriented markers of differentiation. In dysplasias a gradual loss of blood group substances A and B can be observed. Squamous cell carcinomas possess no substances A or B. H antigen as precursor of A and B is increased in preneoplasias and absent in carcinomas. In oral papillomas, leukoplakias, and carcinomas virogen koilocytotic cell changes, papilloma viruses and viral antibodies can be demonstrated. In salivary gland tumors a distinct pattern of distribution for keratin, vimentin, CEA, tissue polypeptide antigen (TPA), metalloproteins, and enzymes can be observed. The cellular stromal reaction (lymphocytes, Langerhans cells, and so forth) can be defined more exactly by monoclonal antibodies.

Topics: Antigens, Viral; Blood Group Antigens; Carcinoembryonic Antigen; Cell Differentiation; Histocytochemistry; Humans; Intermediate Filament Proteins; Keratins; Lactoferrin; Mouth Neoplasms; Peptides; Receptors, Mitogen; Salivary Gland Neoplasms; Tissue Polypeptide Antigen

Histochemical alterations of lectin binding and keratin distribution in experimental carcinomas of the hamster cheek pouch were obtained following cryotreatment. Cryotreated carcinoma cells showed a characteristic reduction in lectin binding and keratin staining shortly following cryosurgery. Tumor tissue, on the 2nd and 3rd days after cryotreatment, displayed destruction and necrosis with almost a complete loss of lectin binding and keratin staining. The remaining neoplastic cells located in the deeper layer showed positive reaction for both lectin binding and keratin, which is indicative of tumor recurrence. Histochemical staining of lectin binding and keratin proteins were useful markers in cryotreated tumor cells to identify either destruction and necrosis or vital activity of neoplastic growth.

Topics: Animals; Carcinoma, Squamous Cell; Cheek; Cricetinae; Cryosurgery; Keratins; Male; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Receptors, Mitogen

Formalin-fixed, paraffin-embedded biopsies of metaplastic keratinized oral mucosa (fibromas and leukoplakias), oral mucosa with epithelial dysplasia and oral squamous cell carcinomas were stained with two monoclonal anti-keratin antibodies (AE1 and AE2). Intense suprabasal staining was seen with AE1 in metaplastic keratinized epithelium, whereas staining of adjacent normal unkeratinized epithelium generally was restricted to basal cells. In dysplastic epithelium and squamous cell carcinomas, staining with AE1 revealed a highly disturbed anti-keratin staining pattern. AE2 stained metaplastic keratinized epithelium in a suprabasal pattern but adjacent unkeratinized epithelium did not stain. In dysplastic epithelium and squamous cell carcinomas, AE2 staining was variable and sometimes absent. Further studies are indicated to clarify whether changes in anti-keratin staining patterns can be used for diagnostic and prognostic purposes.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Epithelium; Erythroplasia; Fibroma; Humans; Keratins; Leukoplakia; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Staining and Labeling

Seventeen cases of verrucous carcinoma of the oral cavity were reviewed. It was found that cytologic features generally associated with viral modification were observed in 15 of these cases. This finding suggests that viruses may play some role in the pathogenesis of verrucous carcinoma. The hypothesis that an opportunistic, persistent virus may act in concert with frank carcinogens to promote the development of verrucous carcinoma is discussed.

Topics: Adult; Aged; Animals; Carcinoma, Papillary; Cell Nucleus; Cell Transformation, Neoplastic; Cell Transformation, Viral; Epithelium; Female; Humans; Hyperplasia; Keratins; Male; Middle Aged; Mouth Neoplasms; Tumor Virus Infections

Seventy-six cases of tumorlike and neoplastic lesions from epidermis and oral epithelium were analyzed by a histochemical technique for the demonstration of keratin. Formalin-fixed paraffin sections were reacted with rabbit antihuman keratin antiserum (dilution of 1:40). The types of distribution of keratin in cells of lesions were classified into five categories: (1) regional, as found in normal squamous epithelia and benign hyperkeratinized lesions, and papilloma, and keratinized squamous cell carcinoma; (2) total, as seen in intensely keratinized lesions, such as verruca vulgaris and highly keratinized squamous cell carcinoma; (3) negative, as displayed by basal cell carcinoma; (4) scattered, as in the most poorly differentiated squamous cell carcinomas; and (5) mixed cellular, as found in both poorly and moderately differentiated squamous cell carcinomas.

Topics: Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Dermatitis, Seborrheic; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Keratosis; Lectins; Mouth Neoplasms; Papilloma; Protein Binding; Skin Neoplasms; Warts

Topics: Carcinoma; Humans; Keratins; Leukoplakia, Oral; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms

The nature of the tumor cells in 5 cases of ameloblastomas was studied by immunohistochemistry, and the findings were compared with developing mouse and human teeth as well as with 5 cases of carcinomas in the oral region. The antigens investigated were keratin, an intracellular cytoskeletal protein typical of epithelial cells, and laminin, an extracellular matrix protein found in basement membranes. Our results show that keratin is expressed by all types of epithelial cells in ameloblastomas as well as in the epidermoid carcinomas, and developing teeth. The epithelial, keratin-positive tumor islands in the ameloblastomas were surrounded by a continuous line of laminin, in a pattern similar to that seen in developing tooth. Laminin was seen also around the epidermoid carcinomas but large areas devoid of laminin were constantly seen between the stroma and the neoplastic epithelium. This indicates a lack of proper basement membrane formation by the malignant epidermoid carcinomas. This may be due either to a diminished production or an increased degradation of basement membrane proteins by the carcinoma cells. Our results are in line with suggestions that ameloblastomas are derived from odontogenic epithelial cells. Immunostaining for keratin does not distinguish between carcinomas and the ameloblastomas. However, visualization of basement membrane proteins such as laminin can apparently be used in the differential diagnosis between ameloblastomas and carcinomas.

Topics: Ameloblastoma; Animals; Basement Membrane; Carcinoma, Squamous Cell; Diagnosis, Differential; Fluorescent Antibody Technique; Histocytochemistry; Immunoenzyme Techniques; Keratins; Laminin; Mice; Mouth Neoplasms; Tooth

Human lymphocytes from a lymph node draining the tumor-bearing area of a patient with a large primary squamous cell carcinoma of the oral mucosa were fused with the nonproducer mouse myeloma, NS-1, to produce interspecies hybridomas. Of 95 hybridoma culture supernatants tested, 23 contained from 0.5 to 50 micrograms/ml of human IgM or IgG. Six supernatant fluids containing greater than 15 micrograms/ml of Ig were tested by indirect immunoperoxidase and immunofluorescence against sections of the autologous carcinoma. Five IgM (lambda) monoclonal antibodies stained the cytoplasm of autologous and allogeneic squamous carcinoma cells. All five monoclonal antibodies stained all layers of normal epidermis but each antibody stained the superficial keratin layer most intensely. Two of the five hybridoma antibodies were further tested. Both antibodies stained all types of normal epithelium; a network of fibers characteristic of intermediate filaments in cultured squamous carcinoma cells and cultured fibroblasts; Z lines in skeletal muscle; and axons in peripheral nerve fibers. We conclude that all five IgM monoclonal antibodies recognize cytokeratins associated with the autologous squamous cell carcinoma. Two of the five hybridoma antibodies recognize an antigenic determinant common to all types of intermediate filament proteins. These data indicate that cytokeratins released by squamous carcinoma cells induced an antibody response in this patient.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Carcinoma, Squamous Cell; Clone Cells; Humans; Hybridomas; Keratins; Lymph Nodes; Male; Mice; Middle Aged; Mouth Neoplasms

Topics: Adult; Carcinoma; Female; Humans; Keratins; Keratosis; Leukoplakia, Oral; Lichen Planus; Male; Middle Aged; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Nicotiana; Plants, Toxic; Precancerous Conditions

Topics: Aged; Candida albicans; Carcinoma, Squamous Cell; Diagnosis, Differential; Epithelium; Glycogen; Humans; Keratins; Leukoplakia, Oral; Lichen Planus; Middle Aged; Mitosis; Mouth Diseases; Mouth Neoplasms

Dyskeratotic cells were examined with light and electron microscopy in human oral leukoplakias and carcinomas and in chemically-induced oral premalignant and malignant lesions of mice and rats. Specific antisera against small and large keratins were used to analyse the distribution of keratin polypeptides. In normal oral mucosa, basal cells did not react with antibodies against large keratins in contrast to the suprabasal cells which did react. Small keratins were found in all epithelial cell layers. In human and experimental premalignant and malignant lesions, intensely labelled dyskeratotic cells were seen, which contained small and large keratins regardless of their position in the epithelium. At the ultrastructural level, these cells showed dense aggregates of tonofilaments. Dyskeratotic cells were often seen in advanced stages of degeneration of phagocytosis of these cells by macrophages and giant cells was frequent. The disturbance of the keratinization process in oral precancer and cancer is easily visualized using keratin antisera. Keratin analysis can give new insights in epithelial maturation and may be helpful for the classification of oral leukoplakia.

Topics: Animals; Female; Humans; Immunoenzyme Techniques; Keratins; Leukoplakia, Oral; Mice; Mice, Inbred Strains; Microscopy, Electron; Mouth Neoplasms; Neoplasms, Experimental; Precancerous Conditions; Rats; Rats, Inbred Strains

Antibodies against different fractions of keratins can be helpful in various fields of special pathology. Antibodies against "small" and "large" keratins permit to evaluate epithelial maturation in skin and oral mucosa. In addition, disturbances of keratinization during inflammatory processes and malignant transformation can be analyzed. The main application of antibodies against the entire fractions of keratins is the detection of the epithelial nature of a neoplasm. By this tool, particular problems in surgical pathology concerning differential diagnosis can be handled in an easier way. Among the different tissues and their neoplasms, examples of the analysis of thymus tumours and salivary gland tumours are presented. Immunoreactivity with keratin antibodies depends crucially on tissue processing. In the normal diagnostic procedure, good results are regularly obtained if cryostat or Bouin-fixed paraffin-embedded sections are used.

Topics: Adenoma; Carcinoma in Situ; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Cytoskeleton; Humans; Keratins; Mouth Mucosa; Mouth Neoplasms; Parakeratosis; Salivary Gland Neoplasms; Salivary Glands; Skin; Skin Neoplasms; Thymus Gland; Thymus Neoplasms

A clinical and histopathologic analysis of 464 oral squamous cell papillomas is presented. Data on age, sex, race, location, clinical appearance, duration, recurrence, and clinical diagnosis are reviewed. One hundred seventy-six of the 464 specimens were examined for hyperkeratosis, character and amount of inflammatory infiltrate, and evidence of cellular atypia. The trends seen in this study support claims made by previous authors regarding incidence and inflammatory involvement. The data support a slightly higher occurrence rate in males than in females and in white as opposed to black patients. Papillomas were most abundant on the palatal complex, dorsum and lateral tongue borders, and lower lips, respectively. Confusion of papilloma for fibroma in the clinical diagnosis was less common than expected. Recurrence rate and incidence of multiple papillomas were low. Histologic study revealed a tendency for hyperkeratotic lesions to arise from nonkeratinized oral sites. Cellular atypia was found, but it is still unclear whether these changes are preneoplastic or due to an increased growth rate.

Topics: Adolescent; Adult; Aged; Child; Child, Preschool; Female; Humans; Hyperplasia; Inflammation; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Recurrence, Local; Papilloma; Time Factors

Topics: Capillaries; Epithelium; Humans; Irritants; Keratins; Mouth Mucosa; Mouth Neoplasms

Topics: Candidiasis, Oral; Carcinoma in Situ; Diagnosis, Differential; Epithelium; Erythroplasia; Humans; Keratins; Keratosis; Leukoedema, Oral; Leukoplakia, Oral; Lichen Planus; Lupus Erythematosus, Discoid; Melanins; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Nevus; Precancerous Conditions; Risk; Sebaceous Glands; Smoking; Stomatitis

During treatment of keratinizing squamous cell carcinomas with bleomycin tumor cells are devitalized by keratinization, while simple necrosis plays a minor role. Connected with this process is a marked resorptive granulomatous inflammation with numerous macrophages which is followed by a fibrous organization. In the border region of the keratinized tumor areas many multinucleated giant cells appear. The nature of these giant cells was the subject of controversy. Enzyme histochemical, electronmicroscopic, and ultrahistochemical investigations in three cases of advanced squamous cell carcinoma of the oral cavity prove that the giant cells which are formed during bleomycin treatment are not multinucleated tumor cells, but multinucleated macrophages. The enzymatic pattern is similar to macrophages with a high content of acid phosphatase and aminopeptidase. The ultrastructure of the giant cells is characterized by lysosomes with acid phosphatase activity, pinocytotic vesicles, and cytoplasmic projections on the cell surface with signs of macroendocytosis. The tumor cells show an epithelial differentiation with desmosomes, tonofibrils, and keratohyaline granula. The giant cells are formed by fusion of mononucleated (monocytogenic) macrophages. The fusions seem to be related to the functional status of the cells. It is possible, that the macrophages and the giant cells have an additional immunologic function. This is suggested by the frequent association of giant cells with lymphocytes. The importance of these facts for the evaluation of the action of bleomycin and the consequences for its therapeutic use are discussed. A combination with methods causing a dedifferentiation of the tumor or suppression of the immunologic defense seems to be problematic.

Topics: Acid Phosphatase; Aged; Bleomycin; Carcinoma, Squamous Cell; Cell Fusion; Cell Membrane; Female; Histocytochemistry; Humans; Keratins; Leucyl Aminopeptidase; Lysosomes; Macrophages; Male; Middle Aged; Mouth Neoplasms; Organoids; Phagocytosis

The keratohyalin granules from 25 human oral leukoplakias, showing benign hyperorthokeratosis histologically, were examined employing a series of histochemical techniques. The tissues were fixed in 10% neutral buffered formalin, 80% methanol, or Carnoy's fluid. The keratohyalin granules stained intensely with Pauly's reagent, Congo red and Harris hematoxylin, indicating the presence of proteins. This was confirmed by abolishing the staining reaction by pretreatment with proteolytic enzymes. The keratohyalin granules also reacted with methyl green-pyronin by staining pink at their peripheries; this staining was abolished by pretreatment with ribonuclease, indicating the presence of ribonucleotides. The keratohyalin granules partially stained with toluidine blue and colloidal iron, indicating the presence of acid polysaccharides. The keratohyalin granules did not react with the Feulgen reagent, suggesting the absence of DNA. Our studies indicate that the keratohyalin granules in human oral leukoplakia are primarily protein(s) complexed with polyribonucleotides. The presence of a carbohydrate moiety suggests the possibility of a protein-polysaccharide component in the granules.

Topics: Adult; Aged; Biopsy; Cytoplasmic Granules; Humans; Hyalin; Keratins; Leukoplakia, Oral; Middle Aged; Mouth Neoplasms; Staining and Labeling

An electron microscope study of tissue diagnosed as white sponge nevus suggested a more advanced process of cellular keratinization in the buccal mucosa than may be appreciated in histologic sections. Ultrastructural features which are normally associated with keratinizing epithelium were observed in the lesion and included (1) a marked increase in the intracellular content of tonofilaments, (2) numerous tonofilament-related keratohyaline granules, (3) membrane-coating granules that possessed an internal striated appearance, (4) the apparent accumulation in the intercellular space of the contents of the striated microgranules, and (5) the presence within some cells of a keratin-like material. Other features included the formation of perinuclear electron-lucent zones and the presence of elongated cytoplasmic processes which produced a complex pattern of interdigitation among the epithelial cells.

Topics: Adult; Cell Nucleus; Cytoplasmic Granules; Desmosomes; Epithelial Cells; Epithelium; Female; Humans; Hyalin; Keratins; Microscopy, Electron; Mouth Mucosa; Mouth Neoplasms; Nevus

A correlative histocytological study for keratinization was done in 446 cases of oral leukoplakia. Cytologically, keratinization could be correctly identified in 91% of leukoplakias exhibiting orthokeratosis, 73% of leukoplakias exhibiting parakeratosis, and 78% of cases showing both types of keratinization. Cytologically, orthokeratosis was present in 83% of individuals with homogeneous leukoplakia and in 69% with ulcerated leukoplakia, while parakeratosis was present in 7% of homogeneous leukoplakias and in 19% of ulcerated leukoplakias. A higher frequency of dysplasia was observed in smears and their corresponding histological sections for those cases which had revealed only parakeratosis in the cytological examination and for both orthokeratosis and parakeratosis as compared to those showing only orthokeratosis. It is suggested that the cytological method is a reliable method for studying oral keratinization and is of value to the clinician in identifying the type of leukoplakias which may need to be biopsied for further surveillance.

Topics: Cytoplasmic Granules; Humans; Keratins; Keratosis; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Parakeratosis

Oral carcinoma in situ (CIS) as a histopathologic entity was studied in seventy-seven patients to determine the clinical and histologic parameters of the disease. There were forty-nine male and twenty-eight female patients, with 45.1 per cent of the lesions being described clinically as white, 15.9 per cent as red, and 8.5 per cent as a combination of the two. The high-risk sites for CIS were floor of the mouth (23.2 per cent of all lesions), tongue (22.0 per cent), and lips (in males only, 19.5 per cent). Histologically, there was a considerable range of variation in surface keratinization, thickness of epithelium, and certain cytologic alterations. The most consistent of all cytologic changes was loss of orientation of cells. There is no information available concerning possible regression of oral CIS, as is known for CIS of uterine cervix. Furthermore, there is no information concerning the frequency of or the period of transition from oral CIS to invasive carcinoma or whether all oral carcinoma is preceded by CIS. Further studies on this disease are essential.

Topics: Adult; Aged; Carcinoma in Situ; Cell Nucleus; Color; Cytoplasm; Epithelial Cells; Female; Humans; Keratins; Lip Neoplasms; Male; Middle Aged; Mouth Floor; Mouth Mucosa; Mouth Neoplasms; Palatal Neoplasms; Precancerous Conditions; Sex Factors; Tongue Neoplasms

Topics: Adolescent; Adult; Female; Humans; Keratins; Male; Mandibular Neoplasms; Mouth Neoplasms; Odontogenic Cysts; Radiography

The occurrence of intracytoplasmic desmosomes in normal, hyperplastic, and hyperkeratotic epithelia, in carcinoma-in-situ and in invasive carcinoma of the human oral cavity is demonstrated by electron microscopy. The mechanism for formation of these structures by invagination, separation and by intracytoplasmic incorporation of plasma membrane-desmosome-complexes are described in various oral epithelia, and other possible mechanisms are discussed. Intracytoplasmic desmosomes may occur in normal and pathological keratinocytes of all layers of the oral epithelium. Their ultrastructure in the peripheral cytoplasm is similar to that of the regular desmosomes on the cell surface. However, as they migrate centripetally, they show signs of degeneration, suggesting dissolution by lysosomal enzyme systems. Various surface membrane alterations involved in the formation of intracytoplasmic desmosomes may lead to a reduction of plasma membrane material and of desmosome structures and to defective intercellular adhesion. The intracytoplasmic incorporation of desmosome structures is a ubiquitous phenomenon exhibited by epithelial keratinocytes under certain physiological or pathological conditions.

Topics: Carcinoma in Situ; Carcinoma, Squamous Cell; Cell Membrane; Cytoplasm; Desmosomes; Epithelial Cells; Epithelium; Humans; Keratins; Mouth Neoplasms; Palate; Tongue; Tongue Neoplasms

Topics: Areca; Biopsy; Histocytochemistry; Humans; Keratins; Mastication; Mouth Mucosa; Mouth Neoplasms; Nicotiana; Plants, Medicinal; Plants, Toxic; Precancerous Conditions; Smoking

Topics: Areca; Carcinoma, Squamous Cell; Humans; Keratins; Mastication; Microscopy, Electron; Mouth Mucosa; Mouth Neoplasms; Nicotiana; Plants, Medicinal; Plants, Toxic; Precancerous Conditions; Smoking

Topics: Adult; Aged; Carcinoma, Squamous Cell; Epithelial Cells; Female; Humans; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Sex Factors

Topics: Diagnosis, Computer-Assisted; Giant Cell Tumors; Humans; Infant; Jaw Neoplasms; Keratins; Leukoplakia, Oral; Lichen Planus; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Odontogenic Cysts; Odontogenic Tumors; Precancerous Conditions; Tooth; Tooth, Deciduous

Topics: Animals; Antibiotics, Antineoplastic; Biopsy; Bleomycin; Carcinoma, Squamous Cell; Culture Techniques; Desmosomes; Ear Neoplasms; Epiglottis; Glycogen; Humans; Keratins; Laryngeal Neoplasms; Maxillary Neoplasms; Mice; Microscopy, Electron; Mouth Neoplasms; Neoplasms, Experimental; Palatal Neoplasms; Tongue Neoplasms; Tonsillar Neoplasms

Topics: Carcinoma, Squamous Cell; Cell Differentiation; Cell Transformation, Neoplastic; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Microtomy; Mouth Neoplasms; Neck Dissection

Topics: Alkaline Phosphatase; Embryo, Mammalian; Fetus; Gestational Age; Glycosaminoglycans; Histocytochemistry; Humans; Keratins; Mouth Mucosa; Mouth Neoplasms

Topics: Adult; Aged; Biopsy; Cytoplasmic Granules; Epithelium; Female; Gingiva; Glycogen; Histocytochemistry; Humans; Keratins; Leukoplakia; Leukoplakia, Oral; Lichen Planus; Male; Microscopy, Electron; Middle Aged; Mouth Diseases; Mouth Mucosa; Mouth Neoplasms; Palate; Tongue

Two hundred and ninety-nine oral leukoplakias were examined by exfoliative cytology; 83 of these lesions were biopsied because of clinical or previous histological features. Epithelial atypia was found in 16 of these biopsies, exfoliative cytology detecting epithelial atypia in only 6 of these cases. Thus exfoliative cytology used alone would have led to a false negative diagnosis in 10 out of 16 (62%) of the cases with epithelial atypia verified by histology. In addition, in no instance was exfoliative cytology responsible for the detection of an epithelial atypia that had been overlooked by consideration of clinical or previous histological features. Exfoliative cytology was successful in the detection of atypia only in those cases in which the surface of the lesion was either ulcerated or not keratinized; all keratinized lesions with atypia yielded negative cytology. The results of this study lead to the conclusion that exfoliative cytology is not to be recommended as a routine diagnostic or screening procedure for the detection of possibly pre-malignant features in oral leukoplakias.

Topics: Biopsy; Cytodiagnosis; Denmark; Epithelium; False Negative Reactions; Humans; Keratins; Leukoplakia; Leukoplakia, Oral; Mass Screening; Mouth Neoplasms

Topics: Cell Differentiation; Cell Division; Culture Techniques; Epithelium; Histocytochemistry; Humans; Keratins; Keratosis; Leukoplakia; Leukoplakia, Oral; Lichen Planus; Metaplasia; Mouth Mucosa; Mouth Neoplasms; Organ Culture Techniques

Topics: Cell Membrane; Cell Nucleus; Cheek; Cytoplasmic Granules; Epithelium; Glycogen; Histocytochemistry; Humans; Inflammation; Keratins; Keratosis; Leukoplakia; Leukoplakia, Oral; Lichen Planus; Microscopy, Electron; Mouth Mucosa; Mouth Neoplasms; Smoking

Sixty-two "leukoplakias" from the cheeks of betel-nut chewers in West Malaysia were studied histologically. Ten biopsies were from non-tobacco betel-nut chewers. An amorphous von Kossa positive layer was seen on the keratin surface in 42 specimens. Tobacco did not appear essential for its formation, and it appeared to be significantly associated with parakeratosis. Its possible significance as a cuticle-like layer prolonging contact between carcinogens and the mucosa is discussed.Parakeratosis appeared to be the most common form of cornification seen, and the mitotic activity in parakeratinized leukoplakias appeared to be significantly greater than orthokeratinized leukoplakias.Comparison with studies on other population samples using different quids suggested that severe histological changes were more likely to be seen when tobacoo-containing quids were chewed as compared to non-tobacco-containing quids.An attempt to correlate the histological changes seen with the clinical habit in leukoplakias from chewers using tobacco-containing quids suggested that epithelial atrophy appeared to be significantly related to the duration of the habit but not to the "intensity" of the habit.

Topics: Adult; Aged; Areca; Biopsy; Connective Tissue; Epithelium; Humans; Keratins; Keratosis; Leukoplakia; Leukoplakia, Oral; Middle Aged; Mitosis; Mouth Mucosa; Mouth Neoplasms; Plants, Medicinal

Topics: Adult; Aged; Carcinoma, Squamous Cell; Esophageal Neoplasms; Female; Granuloma; Humans; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Radium; Skin Diseases; Urinary Bladder Neoplasms; Uterine Cervical Neoplasms

Topics: Adolescent; Child; Humans; Keratins; Leukoplakia; Mouth Mucosa; Mouth Neoplasms; Nevus; Nevus, Pigmented; Pathology; Skin Neoplasms

Topics: Biomedical Research; Blood Chemical Analysis; Carotenoids; Cytodiagnosis; Geriatrics; Humans; Keratins; Leukoplakia; Leukoplakia, Oral; Mouth Neoplasms; Pathology; Pharmacology; Sweden; Toxicology; Vitamin A