cyclin-d1 and Tongue-Neoplasms

Identifying informative prognostic biomarkers for oral tongue squamous cell carcinoma (OTSCC) is of great importance in order to better predict tumour behaviour and to guide treatment planning. Here, we summarise existing evidence regarding immunohistochemical prognostic biomarkers for OTSCC.. A systematic search of the literature was performed using the databases of Scopus, Ovid Medline, Web of Science and Cochrane Library. All studies which had investigated the prognostic significance of immunohistochemical biomarkers in OTSCC during the period from 1985 to 2015 were retrieved. For the five most often evaluated biomarkers a random-effects meta-analysis on overall survival was performed, including those studies that provided the necessary statistical results.. A total of 174 studies conducted during the last three decades were found, and in these 184 biomarkers were evaluated for the prognostication of OTSCC. The five biomarkers most frequently assessed were p53, Ki-67, p16, VEGFs and cyclin D1. In the meta-analyses, the most promising results of the prognostic power for OTSCC were obtained for cyclin D1. For studies of VEGF A and C the results were equivocal, but the pooled analysis of VEGF A separately showed it to be a useful prognosticator for OTSCC. There was no sufficient evidence to support p53, Ki-67 and p16 as prognostic biomarkers for OTSCC. Limitations in the quality of the published studies (e.g., small cohorts, lack of compliance with REMARK guidelines) are widespread.. Numerous biomarkers have been presented as useful prognosticators for OTSCC, but the quality of the conduct and reporting of original studies is overall unsatisfactory which does not allow reliable conclusions. The value of two biomarkers (VEGF-A and cyclin D1) should be validated in a multicentre study setting following REMARK guidelines.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Humans; Ki-67 Antigen; Prognosis; Tongue Neoplasms; Tumor Suppressor Protein p53; Vascular Endothelial Growth Factor A

The chemokine receptor 4 (CXCR4) plays an important role in tumor progression. Overexpressed CXCR4 is associated with a poor prognosis of patient with head and neck squamous cell carcinomas. However, the correlation between CXCR4 and chemotherapy resistance in tongue squamous cell carcinoma (TSCC) remains obscure.. Stable cisplatin-resistant CAL27 CDDP and SCC25 CDDP cells were established and identified by CCK8 assay, and the CXCR4 expression was detected using qRT-PCR and Western blot. CXCR4-siRNA was transfected into TSCC CDDP cells, whose transfect efficiency was examined. Cisplatin sensitivity was further detected, as well as several proliferation and apoptosis-related proteins.. CAL27 CDDP and SCC25 CDDP cells were successfully established, which exhibited significantly higher cell viability and less apoptosis under cisplatin stimulation than that of parental cells. CXCR4 expression was increased in TSCC CDDP cells. After transfection of CXCR4-siRNA, the expression of CXCR4 was reduced by 73% and 78% in CAL27 CDDP and SCC25 CDDP cells, respectively. CCK8 assay and flow cytometry assay revealed that the proliferative capacity under cisplatin stimulation significantly decreased after CXCR4 silencing. Moreover, increased TSCC CDDP cells were arrested in the G0/G1 phase after knockdown of CXCR4. Compared with negative control group, the expression of cyclin D1 and p-AKT decreased, while that of p-caspase-3 and Bax significantly increased.. Silencing CXCR4 may evidently inhibit proliferation, induce apoptosis and enhance cisplatin sensitivity of TSCC CDDP cells by reduced cyclin D1 and p-AKT, and increased p-caspase-3 and Bax.

Topics: Antineoplastic Agents; Apoptosis; bcl-2-Associated X Protein; Carcinoma, Squamous Cell; Caspase 3; Cell Proliferation; Cisplatin; Cyclin D1; Drug Resistance, Neoplasm; Gene Silencing; Humans; Receptors, CXCR4; RNA, Small Interfering; Tongue Neoplasms; Tumor Cells, Cultured

EMS1 (chromosome eleven, band q13, mammary tumor and squamous cell carcinoma-associated gene 1) gene amplification and the concomitant cortactin overexpression have been reported to associate with poor prognosis and tumor metastasis. In this study, we examined cortactin expression by immunohistochemistry in human oral tumors and murine tongue tumors which were induced by the carcinogen 4-nitroquinoline 1-oxide (4-NQO). The immunostaining results show over- to moderate expression of cortactin in 85% (104/122) of oral squamous cell carcinoma (OSCC) tissues and in all 15 leukoplakia tissues examined. Further, statistical analysis indicates that cortactin overexpression appears to be a predictor for shorter survival and poorer prognosis in OSCC patients. In an animal model, cortactin is shown to upregulate in infiltrating squamous cell carcinoma, papilloma, and epithelia with squamous hyperplasia, indicating that cortactin induction is an early event during oral carcinogenesis. It is suggested that cortactin expression is mediated in the progression of pre-malignancy to papilloma, based on earlier cortactin induction in pre-malignancy preceding cyclin D1 in papilloma. In conclusion, cortactin overexpression is frequently observed in human OSCC and mouse tongue tumors. Thus, cortactin may have an important role in the development of oral tumors in human and mice. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 799-812, 2017.

Topics: 4-Nitroquinoline-1-oxide; Adult; Animals; Areca; Carcinogenesis; Carcinoma, Squamous Cell; Cortactin; Cyclin D1; Disease Models, Animal; Disease-Free Survival; Female; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Leukoplakia; Male; Mice; Mice, Inbred BALB C; Middle Aged; Mouth Neoplasms; Plant Extracts; Polymerase Chain Reaction; Prognosis; Proportional Hazards Models; Tongue Neoplasms; Up-Regulation

Accurate nodal staging is pivotal for treatment planning in early (stage I-II) oral cancer. Unfortunately, current imaging modalities lack sensitivity to detect occult nodal metastases. Chromosomal region 11q13, including genes CCND1, Fas-associated death domain (FADD), and CTTN, is often amplified in oral cancer with nodal metastases. However, evidence in predicting occult nodal metastases is limited.. In 158 patients with early tongue and floor of mouth (FOM) squamous cell carcinomas, both CCND1 amplification and cyclin D1, FADD, and cortactin protein expression were correlated with occult nodal metastases.. CCND1 amplification and cyclin D1 expression correlated with occult nodal metastases. Cyclin D1 expression was validated in an independent multicenter cohort, confirming the correlation with occult nodal metastases in early FOM cancers.. Cyclin D1 is a predictive biomarker for occult nodal metastases in early FOM cancers. Prospective research on biopsy material should confirm these results before implementing its use in routine clinical practice. © 2016 Wiley Periodicals, Inc. Head Neck 39: 326-333, 2017.

Topics: Adult; Aged; Aged, 80 and over; Biopsy, Needle; Carcinoma, Squamous Cell; Cortactin; Cyclin D1; Databases, Factual; Early Diagnosis; Fas-Associated Death Domain Protein; Female; Gene Amplification; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Lymphatic Metastasis; Male; Middle Aged; Mouth Floor; Mouth Neoplasms; Netherlands; Observer Variation; Predictive Value of Tests; Prognosis; Reproducibility of Results; Retrospective Studies; ROC Curve; Tongue Neoplasms

This study investigated the effect of RhoA silencing through RNA interference on proliferation and growth of tongue cancer cells, as well as explored the possible mechanisms of this effect.. SSC-4 tongue cancer cells were cultured in vitro and then transfected with small interfering RNA to knock down RhoA expression. The tested cells were divided into three groups: experimental group (experimental group 1: transfected with RhoA-siRNA-1; experi-mental group 2: transfected with RhoA-siRNA-2), negative control group (transfected by random sequence NC-siRNA), and blank control group (transfected with Lipofectamine). The expression levels of RhoA mRNA were respectively measured by quantitative real-time polymerase chain reaction and western blot assay. Moreover, the expression levels of cyclin D1, p21, and p27 and RhoA protein were evaluated by Western blot assay. Proliferation and growth potentiality were analyzed through evaluation of doubling times and methyl thiazolyl tetrazolium assessment.. The expression levels of RhoA gene and protein of experimental groups significantly decreased following siRNA transfection compared with those in the negative and blank control groups. The expression of cyclin D1 decreased significantly and that of p21 and p27 increased significantly. The doubling time was extended and the growth potentiality decreased.. The results indicated that RhoA silencing can inhibit proliferation of tongue cancer cells, whereas RhoA affects cell proliferation by regulating the cell cycle pathway. Thus, RhoA is a potential target in gene therapy for tongue cancer.. 目的 通过RNA干扰技术沉默RhoA基因从而探讨RhoA对舌癌细胞增殖和生长的影响及其作用机制。 方法 体外培养舌鳞状细胞癌SCC-4细胞，以小分子干扰RNA转染沉默RhoA基因的表达。实验分为3组：实验组（又分为实验1组和实验2组，脂质体分别转染对应序列1的RhoA-siRNA和序列2的RhoA-siRNA）、阴性对照组（脂质体转染NC-siRNA）和空白对照组（不转染siRNA）。采用实时定量聚合酶链反应技术检测SCC-4细胞转染后RhoA mRNA的表达，Western blot检测RhoA、Cyclin D1、p21和p27蛋白的表达，四唑盐比色法检测舌癌细胞生长水平和倍增时间。 结果 与阴性对照组和空白对照组相比，实验组舌癌细胞的RhoA基因及蛋白表达降低，p21、p27蛋白表达升高，Cyclin D1蛋白表达降低，细胞倍增时间延长，增殖能力降低（P<0.05）。 结论 沉默RhoA基因可以抑制舌癌细胞的增殖和生长，RhoA基因通过调控细胞周期信号转导途径影响舌癌细胞的增殖，RhoA基因可以成为舌癌基因治疗的靶点。.

Topics: Cell Line, Tumor; Cell Proliferation; Cyclin D1; Gene Silencing; Humans; Neoplasms, Squamous Cell; Real-Time Polymerase Chain Reaction; rhoA GTP-Binding Protein; RNA, Messenger; RNA, Small Interfering; Tongue Neoplasms; Transfection

The typical progression of oral cancer is from hyperplastic epithelial lesions through dysplasia to invasive carcinoma. It is important to investigate malignant oral cancer progression and development in order to determine useful approaches of prevention of dysplastic lesions. The present study aimed to gain insights into the underlying molecular mechanism of oral carcinogenesis by establishing a rat model of oral carcinogenesis using 4‑nitroquinoline 1‑oxide. Subsequently, transcription profile analysis using an integrating microarray was performed. The dynamic gene expression changes of the six stages of rat oral carcinogenesis (normal, mild epithelial dysplasia, moderate dysplasia, severe dysplasia, carcinoma in situ and oral squamous cell carcinomas) were analyzed using component plane presentations (CPP)‑self‑organizing map (SOM). Six genes were verified by quantitative polymerase chain reaction, immunohistochemistry and succinate dehydrogenase (SDH) activity assay kit. Numerous differentially expressed genes (DEGs) were identified during rat oral carcinogenesis. CPP‑SOM determined that these DEGs were primarily enriched during cell cycle, apoptosis, inflammatory response and tricarboxylic acid cycle, indicating the coordinated regulation of molecular networks. In addition, the expression of specific DEGs, such as janus kinase 3, cyclin‑dependent kinase A‑1, B‑cell chronic lymphocytic leukaemia/lymphoma 2‑like 2, nuclear factor‑κB, tumor necrosis factor receptor superfamily member 1A, cyclin D1 and SDH were identified to have high concordance with the results from microarray data. The current study demonstrated that oral carcinogenesis is a multi‑step and multi‑gene process, with a distinct pattern alteration along a continuum of malignant transformation. In addition, this comprehensive investigation provided a theoretical basis for the understanding of the molecular alterations associated with oral carcinogenesis.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinogenesis; Cyclin D1; Disease Models, Animal; Gene Expression Regulation, Neoplastic; Mouth Mucosa; Mouth Neoplasms; Rats; Rats, Wistar; Real-Time Polymerase Chain Reaction; Succinate Dehydrogenase; Tongue Neoplasms; Transcription Factor RelA; Transcriptome

Ectomesenchymal chondromyxoid tumor (ECT) is a rare benign tumor of uncertain lineage, which almost exclusively affects the anterior tongue. Herein, we report 2 cases of ECT occurring in 58- and 56-year-old males on the right and on the left side of the dorsum of the anterior tongue, measuring 18 mm and 10 mm, respectively. Despite positive resection margin in one case, none of the tumors recurred during follow-up of 6 and 5 years. Microscopically, both tumors had lobular architecture with a mixture of solid, microcystic, and chondromyxoid areas. The tumor cells were polygonal or elongated and showed mild atypia in one case. Immunohistochemically, both tumors showed diffuse expression of vimentin and focal positivity of CD10 and of smooth muscle actin. Regarding neural tissue-related markers, there was nearly diffuse expression of CD56 and neuron-specific enolase and focal positivity of PGP 9.5 in both cases and variable expression of CD57, synaptophysin, glial fibrillary acidic protein, and S-100 protein. Interestingly, we observed diffuse expression of SOX10 in one case. In both tumors, diffuse strong nuclear expression of cyclin D1 was present, without CCND1/IGH translocation or CCND1 amplification. The EWSR1 gene rearrangement was not detected. To the best of our knowledge, expression of SOX10, which may support neural crest origin of this peculiar lesion, has not been reported in ECT. The significance of strong cyclin D1 expression remains to be further investigated.

Topics: Biomarkers, Tumor; Cyclin D1; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Male; Mesenchymoma; Microscopy, Electron, Transmission; Middle Aged; Myxoma; Tongue Neoplasms

miR-375 has been implicated in various types of cancers. However, its role in tongue squamous cell carcinoma (TSCC) remains unclear. This study aimed to investigate the effects of miR-375 on cell growth and the prognosis of TSCC patients. Using quantitative reverse transcription-polymerase chain reaction, we evaluated miR-375 expression in TSCC samples and TSCC cell lines. The results showed that miR-375 expression was significantly reduced in the TSCC tissues and cell lines. A low level expression of miR-375 in TSCC patients was related to poor of prognosis. Moreover, the effects of miR-375 overexpression on cell proliferation, the cell cycle and the expression of Sp1 and cyclin D1 were examined in TSCC cells. We demonstrated that overexpression of miR-375 significantly inhibited the cell proliferation and cell cycle progression in TSCC cell lines. Overexpression of miR-375 inhibited Sp1 expression by targeting the 3' untranslated region of the Sp1 transcript. The knockdown of Sp1 expression resulted in the subsequent downregulation of cyclin D1. Taken together, our study suggests that miR-375 inhibits the cell growth, and its expression is correlated with clinical outcomes in TSCC.

Topics: 3' Untranslated Regions; Carcinoma, Squamous Cell; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Down-Regulation; Female; Gene Expression Regulation, Neoplastic; Humans; Male; MicroRNAs; Middle Aged; Prognosis; Sp1 Transcription Factor; Tongue Neoplasms

Thyroid cancer 1 (TC1, C8orf4) plays important roles in many signaling pathways, such as Wnt/β-catenin signaling pathway, and is involved in the development of many cancers. The objective of this study was to examine the expression of TC1 and investigate the associations among TC1, β-catenin, Chibby, cyclin D1, and the clinicopathological factors of oral tongue squamous cell carcinomas (OTSCCs). The expressions of TC1, β-catenin, Chibby, and cyclin D1 were examined in 109 cases of OTSCCs using immunohistochemistry. The expression of TC1 was observed in all cases of OTSCCs but was negative or weak in normal squamous epithelial tissues of tongue. The high expression of TC1 was correlated with the advanced TNM stage (P = 0.042), the abnormal expression of β-catenin (correlation coefficient = 0.314, P = 0.001) and the expression of cyclin D1 (correlation coefficient = 0.274, P = 0.006) in OTSCCs. But we did not find any associations between TC1 and Chibby. The abnormal expression of β-catenin was correlated with the poor differentiation (P = 0.035), advanced TNM stage (P = 0.048) and the expression of cyclin D1 (correlation coefficient = 0.422, P < 0.001). In conclusion, the high expression of TC1 was common in OTSCCs and correlated with the expression of β-catenin and cyclin D1 and the progression of OTSCCs. The high expression level of TC1 might promote the progression of OTSCCs by enhancing the activity of Wnt signaling pathway.

Topics: Adult; Aged; Aged, 80 and over; beta Catenin; Carcinoma, Squamous Cell; Carrier Proteins; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Nuclear Proteins; Tongue Neoplasms; Wnt Signaling Pathway

It is now possible to identify several key factors that determine biological characteristics of squamous cell cancer of the head and neck: genes p53, p16, cyclin D1, P13-K/Akt connected with metastasis proteins (proteases, proteins mesenchymal cells, cell adhesion molecules chemokines), angiogenesis factors (VEGF, PDGF, FGF, TGF-alpha and TGF-beta), IL-8; epidermal growth factor receptors. An important role of tumor cells plays microenvironment. Of course the above mentioned is only a small part of the factors that determine the livelihoods and the activity of cancer cells. All of these factors are potential predictors of the effectiveness of radiation and chemoradiation treatment and actively studied in recent decades.

Topics: Adult; Aged; Aged, 80 and over; Angiogenic Proteins; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoma, Squamous Cell; Chemokines; Chemoradiotherapy; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Gene Expression Regulation, Neoplastic; Humans; Interleukin-8; Male; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Phosphatidylinositol 3-Kinases; Predictive Value of Tests; Prognosis; Prospective Studies; Proto-Oncogene Proteins c-akt; Retrospective Studies; Risk Factors; Tongue Neoplasms; Treatment Outcome; Tumor Suppressor Protein p53

The immunohistochemical expression of Cyclin D1 and Ki-67 were analyzed in tongue squamous cell carcinomas (SCC), relating them to the clinical and morphological exhibition of these tumors.. Twenty-nine patients fulfilled the inclusion criteria; clinical data included gender, age, ethnicity and use of licit drugs such as alcohol and tobacco. The TNM staging and histopathological differentiation grading was assessed for each case. In addition, T1 patients were gathered with T2 patients; and T3 patients were gathered with T4 patients to assemble two distinct groups: (T1/T2) and (T3/T4).. The mean follow-up time was 24 months and 30% of the patients died as a consequence of the disease, while 23.3% lived with the disease and 46.7% lived lesion-free. T1 and T2 tumors showed statistically lesser Ki-67 and Cyclin D1 staining when compared to T3 and T4 tumors.. Ki-67 and Cyclin D1 pose as auxiliary tools when determining the progression of tongue SCC at the time of diagnosis.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Ki-67 Antigen; Male; Middle Aged; Neoplasm Staging; Tongue Neoplasms

Oral squamous cell carcinoma (OSCC) ranks as the fifth most common cancer worldwide with poor prognosis. Recently, tumor necrosis factor receptor-associated factor 4 (TRAF4) has attracted increasing attenuation due to its overexpression in certain cancers. However, its function and underlying mechanism in OSCC remains elusive. In this study, the high expression of TRAF4 mRNA and protein levels was noted in OSCC cell lines. Its overexpression with pcDNA3.1-TRAF4 vector transfection dramatically promoted cell proliferation and inhibited cell apoptosis, indicating a pivotal role of TRAF4 in OSCC cell growth. Simultaneously, TRAF4 elevation also increased cell invasion and migration. Mechanism analysis confirmed that TRAF4 up-regulation induced the expression of β-catenin and the downstream target molecules of cyclinD1, c-myc, Bcl-2, MMP-9 and MMP-2, indicating that TRAF4 could induce the activation of Wnt/β-catenin pathway. After pretreatment with β-catenin siRNA, the pathway was remarkably silenced. Simultaneously, cell growth, invasion and migration induced by TRAF4 were strikingly abrogated, suggesting that TRAF4 may promote OSCC cell growth, invasion and migration by Wnt/β-catenin pathway. Together, this study confirmed that TRAF4 acts as an oncogene for the development and progression of OSCC. Therefore, our study may support a promising therapeutic target for the treatment of OSCC.

Topics: Apoptosis; beta Catenin; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Head and Neck Neoplasms; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Neoplasm Invasiveness; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-myc; RNA Interference; Squamous Cell Carcinoma of Head and Neck; TNF Receptor-Associated Factor 4; Tongue Neoplasms; Transfection; Wnt Signaling Pathway

A small, albeit significant, number of head and neck squamous cell carcinoma (HNSCC) patients has no history of tobacco and alcohol use. Such non-habits associated HNSCCs may represent a distinct clinical entity and exhibit increased aggressiveness. The objective of the study was to understand differences in molecular etiology of habits, and non-habits associated tongue carcinomas.. High-throughput gene expression profiling of 22 tumor samples was carried out. This was followed by quantitative real-time PCR validation of four of the identified differentially expressed genes.. Eighteen genes were identified that correlate strongly with the habits- and non-habits distinction. Among the genes significantly overexpressed in the non-habits group are CCND1, a key cell-cycle regulator, DACT3, a modulator of the Wnt/beta-catenin pathway, and three genes associated with the Notch signaling pathway. CCND1 and DACT3 overexpression in non-habits associated tongue carcinomas were subsequently validated by quantitative real-time PCR in an independent cohort (n = 18) of patient samples. Gene expression data were integrated with publicly available protein interaction data to build a small protein interaction network containing five of 18 differentially expressed genes. This suggested that a functional 'network module' can be implicated in the subgroup distinction. All the tumors analyzed here were human papillomavirus (HPV) negative samples. An association between CCND1 overexpression in oral tumors and poor prognosis has previously been reported. Thus, CCND1 overexpression in non-habits associated anterior tongue carcinomas may contribute to their increased clinical aggressiveness.

Topics: Adaptor Proteins, Signal Transducing; Alcohol Drinking; Alphapapillomavirus; Basic Helix-Loop-Helix Transcription Factors; Carcinoma; Cohort Studies; Cyclin D1; Gene Expression Profiling; Helix-Loop-Helix Motifs; Humans; Prognosis; Protein Interaction Maps; Real-Time Polymerase Chain Reaction; Receptor, Notch1; Receptor, Notch2; Repressor Proteins; Tobacco Use; Tongue Neoplasms; Wnt Signaling Pathway

To investigate the role of interleukin-18 (IL-18) in regulating the growth of the human tongue squamous cell carcinoma cell line CRL-1623™.. The human IL18 gene was cloned and transfected into CRL-1623™ cells using the transfection vector pcDNA3.1(+). Investigations included analysis of cell viability, detection of apoptosis using annexin V-fluorescein isothiocyanate, assessment of caspase 3/7 activity and real-time reverse transcription-polymerase chain reaction to assess expression of the IL18, CCND1 (cyclin D(1)), CCNA1 (cyclin A(1)) and IFNG (interferon-γ) genes.. Introduction of the IL18 gene inhibited cell proliferation at 24, 48 and 72 h after transfection compared with untransfected cells and cells transfected with blank pcDNA3.1(+) vector. Apoptotic cell numbers and caspase 3/7 activity were significantly enhanced by IL18 transfection. Levels of IL18 and IFNG mRNA were elevated and CCND1 mRNA was reduced after 48 h in IL18 transfected cells compared with wild-type cells.. These findings suggest that IL-18 plays a role in the regulation of tongue squamous cell carcinoma and may represent a potential therapeutic target.

Topics: Apoptosis; Carcinoma, Squamous Cell; Caspase 3; Caspase 7; Cell Line, Tumor; Cell Proliferation; Cyclin A1; Cyclin D1; Gene Expression Regulation, Neoplastic; Genetic Vectors; Humans; Interferon-gamma; Interleukin-18; RNA, Messenger; Tongue Neoplasms; Transfection

Several signaling pathways are involved in the progression of squamous cell carcinoma. Among them, activated PI3K/Akt may result in NF-κB nuclear translocation, thus leading to the transcription of genes enrolled in cellular invasion and proliferation, such as cyclin D1. This study sought to evaluate the expression of pAkt, NF-κB and cyclin D1 proteins in head and neck squamous cell carcinoma cell lines and their respective in vitro-obtained invasive clones.. Squamous cell carcinoma cell lines originating from the tongue, pharynx and the metastatic lymph node were submitted to an in vitro invasion assay to select invasive clones. All experimental groups were submitted to immunofluorescence and Western blot assays. Statistical analysis was performed through a Student's t-test with a significance level of 5%.. The pAkt and NF-κB expression differed from cytoplasm and nucleus depending on the studied cell line. The invasive clone from the tongue presented a network-like structure of pAkt's cytoplasmic expression. This lineage as well as the invasive clone from pharynx also showed pAkt and NF-κB nuclear transportation. Significant pAkt and NF-κB increases were observed in the tongue and pharynx invasive clones. Cyclin D1 was detected in the nucleus of all studied cells and was significantly enhanced in the invasive clones from tongue and pharynx.. This study suggests the participation of pAkt, NF-κB and cyclin D1 in the invasion process of head and neck squamous cell carcinoma. Moreover, cytoplasmic pAkt network-like structure was probably related to cytoskeleton changes presented during invasion.

Topics: Active Transport, Cell Nucleus; Carcinoma, Squamous Cell; Cell Line, Tumor; Clone Cells; Cyclin D1; Cytoskeleton; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Humans; Neoplasm Invasiveness; NF-kappa B; Pharyngeal Neoplasms; Proto-Oncogene Proteins c-akt; Signal Transduction; Tongue Neoplasms; Up-Regulation

Several molecular markers have been studied for their usefulness as prognostic markers in oral squamous cell carcinoma (OSCC). One such molecular marker is cyclin D1 which is a proto-oncogene located on 11q13 in humans.. To explore the feasibility of using cyclin D1 as a prognostic marker in tongue and cheek SCC by the fluorescent-in-situ hybridization (FISH) method.. Fifty paraffin-embedded samples (25 each of cheek and tongue SCCs) were obtained from the archives of the Oral Pathology Diagnostic Laboratory. Sociodemographic data, histopathologic diagnoses, lymph node status and survival data were obtained from the Malaysian Oral Cancer Database and Tissue Bank System (MOCDTBS)coordinated by the Oral Cancer Research and Coordinating Centre (OCRCC), University of Malaya. The FISH technique was used to detect the amplification of cyclin D1 using the Vysis protocol. Statistical correlations of cyclin D1 with site and lymph node status were analyzed using the Fisher exact test. Kaplan-Meier and Log Rank (Mantel-Cox) test were used to analyze cyclin D1 amplification and median survival time.. Positive amplification of cyclin D1 was detected in 72% (36) of OSCCs. Detection of positive amplification for cyclin D1 was observed in 88% (22) and 56% (14) of the tongue and cheek tumors, respectively, where the difference was statistically significant (P=0.012). Lymph node metastasis of cheek SCCs showed a trend towards a significant association (P= 0.098) with cyclin D1 amplification whereas the lymph node metastasis of tongue SCC was clearly not significant (P=0.593).There was a statistically significant correlation between cyclin D1 positivity and survival rate (P=0.009) for overall SCC cases and (P<0.001) for cheek SCC cases.. The present study found that cyclin D1 amplification may differ in different subsites of OSCC (tongue vs cheek) and its positive amplification implies an overall poor survival in OSCCs, particularly those arising in cheeks.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cross-Sectional Studies; Cyclin D1; Female; Gene Amplification; Humans; In Situ Hybridization, Fluorescence; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Prognosis; Proto-Oncogene Mas; Survival Rate; Tongue Neoplasms

Oral cancer is a common and lethal malignancy. Direct contact between saliva and the oral cancer lesion makes measurement of tumour markers in saliva an attractive alternative to serum testing.. We tested 19 tongue cancer patients, measuring the levels of 8 salivary markers related to oxidative stress, DNA repair, carcinogenesis, metastasis and cellular proliferation and death.. Five markers increased in cancer patients by 39-246%: carbonyls, lactate dehydrogenase, metalloproteinase-9 (MMP-9), Ki67 and Cyclin D1 (CycD1) (P< or =0.01). Three markers decreased by 16-29%: 8-oxoguanine DNA glycosylase, phosphorylated-Src and mammary serine protease inhibitor (Maspin) (P< or =0.01). Increase in salivary carbonyls was profound (by 246%, P=0.012); alterations in CycD1 (87% increase, P=0.000006) and Maspin (29% decrease, P=0.007) were especially significant. Sensitivity values of these eight analysed markers ranged from 58% to 100%; specificity values ranged from 42% to 100%. Both values were especially high for the CycD1 and Maspin markers, 100% for each value of each marker. These were also high for carbonyls, 90% and 80%, respectively, and for MMP-9, 100% and 79%, respectively.. The significance of each salivary alteration is discussed. As all alterations correlated with each other, they may belong to a single carcinogenetic network. Cancer-related changes in salivary tumour markers may be used as a diagnostic tool for diagnosis, prognosis and post-operative monitoring.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; DNA Glycosylases; Female; Humans; Ki-67 Antigen; Male; Matrix Metalloproteinase 9; Middle Aged; Neoplasm Staging; Saliva; Sensitivity and Specificity; Tongue Neoplasms

To investigate the effect of DAPT (gamma-secretase inhibitor) on the growth of human tongue carcinoma cells and to determine the molecular mechanism to enable the potential application of DAPT to the treatment of tongue carcinoma.. Human tongue carcinoma Tca8113 cells were cultured with DAPT. Cell growth was determined using Indigotic Reduction method. The cell cycle and apoptosis were analyzed by flow cytometry. Real-time PCR and Immuno-Fluorescence (IF) were employed to determine the intracellular expression levels.. DAPT inhibited the growth of human tongue carcinoma Tca8113 cells by inducing G0-G1 cell cycle arrest and apoptosis. The mRNA levels of Hairy/Enhancer of Split-1 (Hes-1), a target of Notch activation, were reduced by DAPT in a dose-dependent manner. Coincident with this observation, DAPT induced a dose-dependent promotion of constitutive Caspase-3 in Tca8113 cells.. DAPT may have a therapeutic value for human tongue carcinoma. Moreover, the effects of DAPT in tumor inhibition may arise partly via the modulation of Notch-1 and Caspase-3.

Topics: Amyloid Precursor Protein Secretases; Antineoplastic Agents; Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Carcinoma; Caspase 3; Cell Line, Tumor; Cell Membrane; Cell Nucleus; Cyclin D1; Dipeptides; Dose-Response Relationship, Drug; G1 Phase; Homeodomain Proteins; Humans; Receptor, Notch1; Repressor Proteins; Resting Phase, Cell Cycle; Tongue Neoplasms; Transcription Factor HES-1

To investigate the expression of JAK, ERK and Cyclin D proteins in squamous-cell carcinoma of tongue.. The expression of JAK, ERK and Cyclin D1 proteins was determined with SP immunohistochemical method in 30 cases of lingual Squamous cell carcinoma, 20 of normal lingual mucosa, 10 of mild epithelial dysplasia and 20 of severe epithelial dysplasia.. The expression of pJAK in lingual squamous-cell carcinoma and epithelial dysplasia was stronger than that of normal lingual mucosa (chi2=37.54, P<0.01), and the expression of pJAK in lingual squamous-cell carcinoma was significantly higher than that of the epithelial dysplasia (chi2=6.83, P<0.05). pJAK expression in squamous-cell carcinoma of low-middle differentiation was stronger than that of high differentiation. There was no significant difference in pERK expression among lingual squamous-cell carcinoma, normal lingual mucosa and epithelial dysplasia. There was a significantly positive correlation between pJAK and Cyclin D1 expression in SCC (r=0.619, P<0.05). There was no significant correlation between the expression of pERK and Cyclin D1 (r=0.231, P>0.05).. Over-expression of pJAK and Cyclin D1 may be associated with the occurrence and development of squamous-cell carcinoma of the tongue.

Topics: Carcinoma, Squamous Cell; Cyclin D1; Extracellular Signal-Regulated MAP Kinases; Humans; Immunohistochemistry; Janus Kinases; Phosphorylation; Tongue Neoplasms

We report an immunohistochemical investigation of the expression of activated extracellular signal-regulated kinase (ERK1/2) and cyclin D1 protein in both oral tongue squamous cell carcinomas (OTSCCs) and normal tongue epithelium. The expression of Ki-67 labeling index (LI) was also examined in order to evaluate cell proliferation activity. The expression of activated ERK1/2, cyclin D1 protein and Ki-67 LI were significantly stronger in OTSCCs than in normal oral mucosa (P<0.05). Both over-expression of activated ERK1/2 and positive expression of Ki-67 in OTSCCs were significantly associated with a moderately or poorly differentiated grade of carcinoma (P<0.05). Cyclin D1 immunostaining showed statistically significant association with both lymph node metastasis (P<0.05) and a tumor thickness >5mm (P<0.05). Over-expression of activated ERK1/2 was positively correlated with cyclin D1 protein expression (P<0.05, r=0.624) and cell proliferation-related indexes Ki-67 (P<0.05, r=0.723). Our results suggest that over-expression of activated ERK1/2 and cyclin D1 protein are involved in oral tongue carcinogenesis, and that activation of ERK1/2 might be related to cell cycle regulation and cell proliferation in OTSCCs.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cell Proliferation; Cyclin D1; Extracellular Signal-Regulated MAP Kinases; Female; Humans; Japan; Ki-67 Antigen; Lymphatic Metastasis; Male; Middle Aged; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Neoplasm Proteins; Tongue Neoplasms

To interfere in the Tca8113-CDDP cell line with siRNA of cyclin D1 and to investigate time and dose dependent gene silencing effect of siRNA of cyclin D1.. siRNA of cyclin D1 was transfected into Tca8113-CDDP cells Fluorescent CY3 dye labeled siRNA GAPDH was used as the control. The transient transfecting efficiency was examined at 4, 24, 48 and 72 h. The relative quantity of the target RNA of cyclin D1 was analyzed with SYBR Green fluorescent dye kit by the Real-time PCR assay. The protein level of cyclin D1 was examined with Western blot. The changes of cisplatin sensitivity after treatment with siRNA cyclin D1 were examined with methyl thiazolyl tetrazolium (MTT) assay.. The optimized transfecting efficiency with CY3 labeled siRNA GAPDH in Tca8113-CDDP cells was over 90%. The silencing rate of cyclin D1 siRNA was 81.6% at 24 h, 80.7% at 48 h and 94.3% at 72 h. Dose-dependent manner of gene silencing effect was observed when the siRNA concentration was lower than 100 nmol/L, however, gene silencing effect reached its platform when the concentration was higher than 100 nmol/L. The protein levels of cyclin D1 at 24, 48 and 72 h after transfection decreased significantly, and so did the growth of cells. Inhibition rates of cell growth induced by cisplatin after administration with or without cyclin D1 siRNA were 58.4% and 34.8%, respectively.. Chemical synthesized cyclin D1 siRNA effectively silenced the expression of cyclin D1 gene in Tca8113-CDDP cells in vitro, with a time- and dose-dependent manner and target gene silence in cells increased its sensitivity to cisplatin.

Topics: Antineoplastic Agents; Carcinoma, Squamous Cell; Cell Line, Tumor; Cisplatin; Cyclin D1; Dose-Response Relationship, Drug; Drug Resistance, Neoplasm; Gene Expression Regulation, Neoplastic; Gene Silencing; Humans; RNA, Small Interfering; Tongue Neoplasms; Transfection

The effects of dietary feeding with a polyisoprenylated benzophenone, garcinol, isolated from Garcinia indica fruit rind on the development of 4-nitroquinoline 1-oxide (4-NQO)-induced oral carcinogenesis were investigated in male F344 rats. At 7 weeks of age, animals were given 4-NQO at 20 ppm in the drinking water for 8 weeks to induce tongue neoplasms. They also received the diets containing 100 or 500 ppm garcinol either during (for 10 weeks) or after (for 22 weeks) the carcinogen exposure. The other rats were given tap water without 4-NQO throughout the experiment, and fed garcinol (500 ppm)-containing diet or basal diet alone. At the end of the study (week 32), incidences of tongue neoplasms and preneoplastic lesions, cell proliferation activity in the normal-like tongue epithelium estimated by 5-bromodeoxyurideine (BrdU)-labeling index and cyclin D1-positive cell ratio, and immunohistochemical expression of cyclooxygenase-2 (COX-2) in the tongue lesions were determined. Dietary garcinol significantly decreased the incidence and multiplicity of 4-NQO-induced tongue neoplasms and/or preneoplasms as compared to the control diet. Dietary administration of garcinol also significantly reduced the BrdU-labeling index and cyclin D1-positive cell ratio, suggesting reduction in cell proliferation activity in the tongue by garcinol. The COX-2 expression in the tongue lesions was also suppressed by feeding with garcinol. These results indicate that dietary administration of garcinol inhibited 4-NQO-induced tongue carcinogenesis through suppression of increased cell proliferation activity in the target tissues and/or COX-2 expression in the tongue lesions.

Topics: 4-Nitroquinoline-1-oxide; Animals; Anticarcinogenic Agents; Carcinogens; Cyclin D1; Cyclooxygenase 2; Diet; Male; Precancerous Conditions; Prostaglandin-Endoperoxide Synthases; Rats; Rats, Inbred F344; Terpenes; Tongue Neoplasms

The INK4A-ARF locus at chromosome 9p21 is frequently altered in head and neck squamous cell carcinoma (SCC) and encodes two distinct tumor suppressors, p16(INK4A) and p14(ARF). This study addressed the role of p14(ARF) as a potential prognostic marker in this disease.. p14(ARF) protein expression was assessed by immunohistochemistry in a cohort of 140 patients with SCC of the anterior tongue. Using univariate and multivariate Cox's proportional hazards models, the outcomes examined were time to disease recurrence or death, with or without clinicopathologic covariates, including nodal status, disease stage, treatment status, Ki-67 staining, and molecular markers with known functional or genetic relationships with p14(ARF) (p16(INK4A), p53, pRb, p21(WAF1/CIP1), E2F-1).. On multivariate analysis, p14(ARF) positivity (nucleolar p14(ARF) staining and/or nuclear p14(ARF) staining in >/=30% of tumor cells) was an independent predictor of improved disease-free survival (DFS; P = 0.002) and overall survival (OS; P = 0.002). This was further enhanced when p14(ARF) positivity was cosegregated with positive (>/=1%) p16(INK4A) staining (DFS, P < 0.001; OS, P < 0.001). Patients whose cancers were p14(ARF) negative and p53 positive (>50%) had the poorest outcome (DFS, P < 0.001; OS, P < 0.001) of any patient subgroup analyzed.. These data show that in patients with SCC of the tongue, combined nuclear and nucleolar expression of p14(ARF) protein predicts for improved DFS and OS independent of established prognostic markers.

Topics: Carcinoma, Squamous Cell; Cell Cycle Proteins; Cohort Studies; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; DNA-Binding Proteins; E2F Transcription Factors; E2F1 Transcription Factor; Humans; Immunohistochemistry; Ki-67 Antigen; Multivariate Analysis; Neoplasm Recurrence, Local; Neoplasm Staging; Predictive Value of Tests; Prognosis; Proportional Hazards Models; Retinoblastoma Protein; Survival Analysis; Tongue Neoplasms; Transcription Factors; Tumor Suppressor Protein p14ARF; Tumor Suppressor Protein p53

Some oral cancers are known to develop from dysplastic oral epithelium. In the present study, the expression of c-Jun, c-Fos, and cyclin D1 proteins in oral epithelial lesions with different degrees of dysplasia, and in oral squamous cell carcinomas (OSCCs) was evaluated. Eighteen cases of mild dysplasia, 23 cases of moderate to severe dysplasia and 24 OSCCs were studied immunohistochemically. Additionally, 15 sections of oral mucosa without any evidence of dysplasia were included in the study.. c-Jun expression increased according to the degree of oral dysplasia, with the greatest expression found in OSCC. c-Fos expression was intense in normal mucosa, reduced in mild dysplasia and high in moderate to severe dysplasia and in OSCCs. Cyclin D1 was expressed in only a few cases of moderate to severe dysplasia and in most of the OSCCs. Statistical analysis showed a correlation between the three proteins and the degree of epithelial alteration. The present results indicate a possible role of c-Jun and c-Fos in malignant transformation of oral mucosa.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Cyclin D1; Humans; Immunohistochemistry; Leukoplakia, Oral; Mouth Mucosa; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-jun; Tongue Neoplasms; Transcription Factor AP-1

Expression of a panel of biomarkers, such as p53, Bcl-2, Cyclin D1, c-myc, p21ras, c-erb B2, cytokeratin-19 (CK-19), and factor VIII-related antigen (FVIII-RA), was studied together in anterior tongue tumors from the oral cavity and in posterior tongue tumors from the oropharynx of patients with early- and locally advanced-stage disease, to evaluate their prognostic value.. The expression of the above-mentioned biomarkers was studied by immunohistochemical localization.. In this study, 18%, 26%, 62%, 75%, 73%, 50%, and 29% of the tumors exhibited p53, Bcl-2, Cyclin D1, c-myc, p21ras, c-erb B2, and CK-19 expression, respectively. Twenty percent of the tumors had a microvessel count of >0.0. The expression of these biomarkers was also correlated with clinicopathologic parameters. In early-stage patients with a tobacco habit, who showed borderline significance for relapse-free survival by Kaplan-Meier survival analysis, this turned out to be significant, with the general linear model univariate survival analysis. In the total group, disease stage emerged as the most significant prognostic factor, followed by c-myc, when Cox forward stepwise regression and general linear model multivariate survival analysis were performed. However, Cyclin D1, which was significant by Cox forward stepwise regression analysis, lost its significance by general linear model multivariate analysis. In patients with early-stage disease, MVC, which was a significant predictor of disease relapse by Cox forward stepwise regression analysis, lost its significance by general linear model analysis because of small number of patients. In patients with locally advanced tongue cancer, multivariate survival analysis of individual biomarkers by both Cox forward stepwise regression and general linear model analysis indicated c-myc expression to be strongly indicative of poor prognosis. However, multivariate analysis of individual markers along with a combination of markers showed that only by Cox forward stepwise regression analysis did the combined expression of markers c-myc, Cyclin D1, and p21ras emerge as a significant independent prognosticator.. Overall stage emerged as the most significant prognostic indicator of disease outcome. Tobacco habit also affected relapse-free survival in patients with early-stage disease. However, immunostaining of c-myc in the tumors of locally advanced-stage tongue cancer patients might be a potential adjunct to clinical stage in the pathologic evaluation of tongue specimens.

Topics: Adult; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Male; Multivariate Analysis; Neoplasm Staging; Prognosis; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins p21(ras); Regression Analysis; Survival Rate; Tongue Neoplasms; Tumor Suppressor Protein p53

Ligands for peroxisome proliferator-activated receptor (PPAR) gamma have been implicated in growth inhibition and cell differentiation in certain malignancies. In this study, the effects of troglitazone, a PPAR gamma ligand, given during the postinitiation phase of oral carcinogenesis initiated with 4-nitroquinoline 1-oxide (4-NQO) were investigated in male F344 rats. Rats aged 6 weeks were given 4-NQO at 20 ppm for 8 weeks to induce tongue neoplasms. Starting 1 week after the cessation of 4-NQO exposure, animals were fed diets containing 0, 30 or 100 ppm troglitazone for 22 weeks. At the end of the study (week 32), the incidences of 4-NQO-induced tongue neoplasms and preneoplasms were determined histopathologically and cell proliferation activity was estimated by counting bromodeoxyuridine (BrdU)-labeling indices and cyclin D1-positive cell ratios. In addition, immunohistochemical expression of cyclooxygenase (COX)-2 and PPAR gamma was assessed in the tongue lesions. Feeding with 100 ppm troglitazone significantly decreased the incidence of squamous cell carcinoma when compared to the group without troglitazone treatment (5.0% vs. 45.8%, P < 0.005). Interestingly, the BrdU-labeling index and cyclin D1-positive cell ratio assessed in the non-lesional tongue squamous epithelium were reduced by dietary administration of troglitazone (P < 0.0001-0.005). Additionally, the immunoreactivity of COX-2 in the tongue lesions was also decreased by the treatment (P < 0.01-0.05). These results clearly showed that dietary troglitazone inhibits 4-NQO-induced tongue carcinogenesis and such inhibition is related to suppression of increased cell proliferation and/or COX-2 expression. This study warrants further investigation on the use of PPAR gamma ligands as a novel preventive approach for oral malignancy.

Topics: 4-Nitroquinoline-1-oxide; Animals; Antineoplastic Agents; Bromodeoxyuridine; Carcinogens; Chromans; Cyclin D1; Cyclooxygenase 2; DNA-Binding Proteins; Incidence; Isoenzymes; Ligands; Male; Precancerous Conditions; Prostaglandin-Endoperoxide Synthases; Rats; Rats, Inbred F344; Receptors, Cytoplasmic and Nuclear; Thiazolidinediones; Tongue; Tongue Neoplasms; Transcription Factors; Troglitazone; Tumor Cells, Cultured

Overexpression of E2F-1 is associated with increased invasiveness in head and neck squamous cell carcinoma cell lines in vitro, but its significance in vivo is unknown. This study sought to determine the relationship between E2F-1 and retinoblastoma protein (pRb) expression and disease outcome in squamous cell carcinoma (SCC) of the anterior tongue.. pRb and E2F-1 protein expression was assessed by immunohistochemistry in a cohort of 145 patients with SCC of the anterior tongue. The outcomes examined were time to disease recurrence or death. The relationships between E2F-1 or pRb expression and outcome were assessed by univariate and multivariate Cox's proportional hazards model, with or without clinicopathological covariates, including nodal status, disease stage, treatment status, and molecular markers (cyclin D1, p16(INK4A), and Ki-67) previously measured in this cohort.. On univariate analysis, increased expression of E2F-1 (>35% of positive-stained nuclei) was associated with increased disease-free survival (DFS; hazard ratio [HR]: 0.35; P = 0.04) and increased overall survival (OS; HR: 0.33; P = 0.06). Decreased expression of pRb (<50% positive nuclei) was associated with increased DFS (HR: 1.81; P = 0.06) but not with OS (P = 0.11). However, when considered simultaneously with other significant factors, i.e. lymph node status, p16(INK4A) protein expression, and histopathological grade, in the multivariate Cox's proportional hazards model, the additional contributions of E2F-1 and/or pRb expression to DFS and OS were not statistically significant.. These data demonstrate that in patients with SCC of the tongue, overexpression of E2F-1 is associated with increased DFS and OS. However, this association is not independent of lymph node status, tumor grade, and p16(INK4A) expression. Among the cell cycle-regulatory molecules studied, p16(INK4A) expression is the most predictive molecular marker of disease outcome.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cell Line, Tumor; Cohort Studies; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Disease-Free Survival; DNA-Binding Proteins; E2F Transcription Factors; E2F1 Transcription Factor; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Male; Middle Aged; Multivariate Analysis; Prognosis; Proportional Hazards Models; Retinoblastoma Protein; Time Factors; Tongue Neoplasms; Transcription Factors; Treatment Outcome

Forty-two specimens from oropharyngeal (tonsil and base of tongue) squamous cell carcinoma patients (SCC) were studied for presence of HPV 16 by in situ hybridization and by immunohistochemistry for p53 and Cyclin D1 protein overexpression. Thirty-one per cent of cases were HPV-16 positive, which correlates with the prevalence reported worldwide. 74% of cases showed p53 protein overexpression and 79% showed Cyclin D1 overexpression. There was no correlation between HPV status and either p53 or Cyclin D1 overexpression (P>0.05). These three variables also did not correlate with factors such as grade of the tumour, stage of the disease or lymph nodal metastasis at presentation.

Topics: Carcinoma, Squamous Cell; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Male; Oncogene Proteins, Viral; Oropharyngeal Neoplasms; Papillomaviridae; Tongue Neoplasms; Tonsillar Neoplasms; Tumor Suppressor Protein p53

The status of cyclin D1 and glycogen synthase kinase 3beta (GSK-3beta) was investigated in 41 patients with T1 and T2 squamous cell carcinomas (SCCs) of the tongue. Out of the 41 SCCs, 27 (65.9%) showed overexpression of cyclin D1 in comparison with normal lingual epithelia by an immunohistochemical method. Cyclin D1 gene amplification was detected in only two (9.1%) of 22 informative cases of the SCCs by differential PCR. Expression of GSK-3beta, which was found to regulate proteosomal degradation of cyclin D1 protein, was reduced in 16 cases (39.0%) of the SCCs relative to normal epithelia, and the intensity of GSK-3beta staining showed an inverse association with cyclin D1. These findings suggest that overexpression of cyclin D1 primarily results from stabilization due to reduction of GSK-3beta, but not cyclin D1 gene amplification, in lingual SCCs. Kaplan-Meier analysis demonstrated that the patients with high cyclin D1 and reduced GSK-3beta expression had a significantly lower 5-year survival than the patients with low cyclin D1 and non-reduced GSK-3beta expression (P=0.014). The cyclin D1 and GSK-3beta coupled assessment was more valuable for the prediction of prognosis than assessment based on cyclin D1.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Female; Follow-Up Studies; Gene Amplification; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Immunoenzyme Techniques; Male; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Polymerase Chain Reaction; Prognosis; Survival Rate; Tongue Neoplasms

The precise mechanism responsible for the frequent overexpression of cyclinD1 in human head and neck squamous cell carcinoma (HNSCC) is not known. In view of the fact that signal transducers and activators of transcription 3 (Stat3) is often activated in HNSCC cells, we examined the effects of Stat3 on cyclin D1 expression and cell proliferation in the YCU-H891 HNSCC cell line that displays constitutive activation of Stat3. Expression of a dominant negative Stat3 construct in YCU-H891 cells inhibited proliferation, cyclin D1 promoter activity, and cellular levels of cyclin D1 mRNA and protein. The levels of the antiapoptotic Bcl-2 and Bcl-X(L) proteins were also inhibited. In 51 primary tumor samples from patients with squamous cell carcinoma of the p.o. tongue, there was a significant correlation between increased levels of the activated form of Stat3, phosphorylated-Stat3, and increased levels of cyclin D1 (P < 0.0001). Increased tumor levels of phosphorylated-Stat3 were also associated with lower survival rates (P < 0.01). This study provides the first evidence that in HNSCC, constitutive activation of Stat3 plays a causative role in overexpression of cyclin D1, and in clinical studies, Stat3 activation may provide a novel prognostic factor. Furthermore, agents that target Stat3 may be useful in the treatment of HNSCC.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Division; Cyclin D1; DNA-Binding Proteins; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Neoplasm Metastasis; Neoplasm Staging; Prognosis; RNA, Messenger; STAT3 Transcription Factor; Tongue Neoplasms; Trans-Activators; Tumor Cells, Cultured

Carcinosarcoma of the tongue is a rare malignancy and its molecular aspect is unclear. A case of carcinosarcoma of the tongue in a 51-year-old man is presented. A polypoid tumor of the tongue, measuring 12 x 12 x 6 mm, was resected. Histologically, the tumor was composed of a squamous cell carcinoma and a spindle cell sarcomatous component. We previously showed that one cell-cycle regulator, the cyclin D1 gene, was frequently amplified in esophageal carcinosarcoma, which shows the same morphologic features as carcinosarcoma of the tongue. In this case, we examined whether the cyclin D1 gene is amplified in carcinosarcoma of the tongue as well. Fluorescence in situ hybridization analysis revealed that the cyclin D1 gene was amplified in both components of carcinosarcoma of the tongue. The cyclin D1 protein was also detected by immunostaining in both components. Our results suggest that the amplification of cyclin D1 gene plays a role in the molecular pathogenesis of carcinosarcoma of the tongue, at least in some cases.

Topics: Carcinosarcoma; Cyclin D1; Gene Amplification; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Male; Middle Aged; Tongue Neoplasms

Cyclin D1 is a cell cycle regulatory factor that modulates a critical step in cell cycle control. Cyclin D1 is overexpressed in a significant proportion of head and neck cancers and correlates with a poor prognosis. Abrogation of cyclin D1 action through antisense cyclin D1 shows promise as an antitumor therapy, with an inhibitory effect in head and neck squamous cell carcinoma both in vitro and in vivo. The suppressive effect of antisense cyclin D1 in head and neck cancer xenografts in nude mice is incomplete, however, suggesting that combination with another antitumor agent is necessary for complete tumor eradication. Cisplatin is a widely used chemotherapeutic agent in head and neck cancer, and is particularly effective in combination with radiation therapy. In this study, we investigate whether antisense cyclin D1 enhances the sensitivity of head and neck cancer cells to cisplatin. Such an enhancement of sensitivity would suggest that combination therapy using antisense cyclin D1 and cisplatin would be an effective treatment modality for head and neck cancer.. Antisense cyclin D1 was transfected into the head and neck squamous cell carcinoma cell line CCL23 using a plasmid vector. Both the parental CCL23 cells and the antisense cyclin D1-transfected CCL23 cells (CCL23AS) were treated with cisplatin at increasing concentrations. The dosage of cisplatin ranged from 1 microg/mL to 10 microg/mL. Initial exposure to cisplatin was for 2 hours, with increasing exposure times in succeeding experiments. Cell viability assays were done following cisplatin exposure. Dose response curves for the two cell lines were plotted and compared. Western blot analyses were done on the cisplatin-treated cell lines to determine levels of cyclin D1 expression.. Increasing concentrations of cisplatin resulted in significantly higher rates of cell killing in the antisense cyclin D1-transfected cells than in the parental cells. The ID50 values for the parental CCL23 cells and the antisense cyclin D1-transfected CCL23 cells were 7 microg/mL and 3 microg/mL, respectively, indicating significant enhancement of sensitivity to cisplatin in the antisense cyclin D1-transfected cells. Western blot analyses demonstrated decreased expression of cyclin D1 in the CCL23AS cells with increasing doses of cisplatin, compared with the parental CCL23 cells.. Antisense cyclin D1-transfected CCL23 cells demonstrate an enhanced sensitivity to the effects of cisplatin compared with the parental cell line. Although the mechanism for this phenomenon is not completely understood, the data suggests the potential use of combination therapy using antisense cyclin D1 and cisplatin for head and neck cancers. While neither agent alone can completely eradicate head and neck cancers, the synergistic effect of the two may be an effective therapeutic protocol for refractory head and neck cancers. Future investigation into the combination of antisense cyclin D1 with cisplatin for treatment of head and neck cancer is needed.

Topics: Carcinoma, Squamous Cell; Cell Survival; Cisplatin; Cyclin D1; Dose-Response Relationship, Drug; Drug Synergism; Gene Expression Regulation, Neoplastic; Humans; Laryngeal Neoplasms; RNA, Antisense; Tongue Neoplasms; Transfection; Tumor Cells, Cultured

We investigated the immunohistochemical expression of Rb protein (pRb), which plays an important role in the regulation of the cell cycle, in rat tongue carcinoma induced by 4-nitroquinoline 1-oxide. In addition, we made an immunohistochemical investigation of cyclin D1 and cdk4, which are involved in the Rb pathway. The labeling index of pRb expression in cases with carcinoma was significantly decreased compared with that in cases with a premalignant lesion (P<0.01), while the labeling index of cyclin D1 and cdk4 increased gradually during the course of carcinogenesis. We analyzed the phosphorylation of pRb by immunoblotting using G3-245 monoclonal antibody, which recognizes both the phosphorylated and unphosphorylated forms of pRb. Although expression of the phosphorylated pRb band was notably increased in dysplastic membrane compared with the control membrane, it almost disappeared in cases with carcinoma. Unphosphorylated pRb bands were also expressed in control membrane and dysplastic membrane but not in cases with carcinoma. In conclusion, a decrease of pRb and an increase of cdk4 and cyclin D1 were shown to occur during the premalignant stage. The decrease of pRb in quantity and the increase of its phosphorylation may prevent G1 arrest and consequently accelerate proliferation of the chemically injured cells contributing to the initiation of carcinogenesis.

Topics: 4-Nitroquinoline-1-oxide; Animals; Blotting, Western; Carcinogens; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinases; Disease Progression; Immunoenzyme Techniques; Male; Neoplasm Proteins; Proliferating Cell Nuclear Antigen; Proto-Oncogene Proteins; Rats; Rats, Sprague-Dawley; Retinoblastoma Protein; Tongue Neoplasms

Amplification of CCND1 was studied in 23 tongue carcinoma patients by fluorescence in situ hybridization (FISH) using a paraffin embedded specimen. All the patients received complete resection of the primary site with or without neck dissection. CCND1 amplification was positive in 13 (56.5%) out of 23 cases. Correlations between CCND1 amplification and histological grading, T category, N category, and Stages were not significant. The 5-year disease-free survival rate, which was 23.1% for CCND1 amplification positive patients and 80.0% for negative patients, was significantly better for the CCND1 amplification negative patients (P=0.0070). Nine patients were examined by dual-color FISH with the probe for centromere of chromosome 11 and 11q13. In five patients, who had positive amplification for CCND1, cell numbers with a larger number of signals for 11q13 than the centromere of chromosome 11 were significantly higher than those of CCND1 amplification negative patients (P=0.013). This indicates that amplification of 11q13 occurs more frequently than aberration of chromosome 11 in CCND1 amplification positive patients. From these results, the amplification of CCND1 is a key factor in predicting the aggressiveness of tongue cancer. Furthermore, FISH proved to be a useful method for such evaluation.

Topics: Carcinoma, Squamous Cell; Chromosomes, Human, Pair 11; Cohort Studies; Cyclin D1; Female; Gene Amplification; Humans; In Situ Hybridization, Fluorescence; Male; Middle Aged; Neoplasm Recurrence, Local; Prognosis; Retrospective Studies; Survival Rate; Tongue Neoplasms

Tongue squamous cell carcinoma makes up a large percentage of head and neck cancers, and the incidence among young patients is increasing. The aim of this study was to reveal the correlation between cyclin D1 (CCND1) expression and clinical and histologic features. We performed an immunohistochemical study on the level of CCND1 expression in tumor specimens obtained from 94 patients with tongue squamous cell carcinoma. The relationship between the expression and the following features such as age, sex, smoking and alcohol intake history, T, N, histologic grade, and multiple primary cancer was analyzed. Eighteen patients (19%) showed CCND1 overexpression (tumor cell nuclei positivity >/=50%). The 5-year survival rate of high CCND1 expressors was 39%, which was significantly poor (p=0.04). N classification correlated with CCND1 expression. CCND1 overexpression is associated with poor survival associated with progression of lymph node spread in patients with tongue squamous cell carcinomas. CCND1 expression may be a useful biologic marker for prognosis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Alcohol Drinking; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Immunohistochemistry; Japan; Male; Middle Aged; Neoplasm Proteins; Prognosis; Smoking; Survival Analysis; Tongue Neoplasms

Cyclin D1 and p16INK4A are molecules with pivotal roles in cell cycle control and the development of diverse human cancers, and overexpression of cyclin D1 and loss of p16INK4A expression are common genetic events in head and neck squamous cell carcinoma. The prognostic significance of these molecular events at different sites within the head and neck, however, remains controversial. Thus, we sought to determine the relationship between cyclin D1 and/or p16INK4A expression and disease outcome in squamous cell carcinoma of the anterior tongue. Immunohistochemical detection of nuclear proteins cyclin D1, p53, and p16INK4A, and the Ki-67 labeling index was undertaken in tissue sections from 148 tongue cancers treated by surgical resection. Nuclear antigen status was analyzed in relation to pathological variables, tumor recurrence, and patient survival. Statistical significance was assessed using chi2 analysis for pathological variables and the Kaplan-Meier method, log rank test, and the Cox proportional hazards model for survival parameters. Overexpression of cyclin D1 occurred in 68% of tumors (100 of 147) and was associated with increased lymph node stage (P = 0.014), increased tumor grade (P = 0.003), and reduced disease-free (P = 0.006) and overall (P = 0.01) survival. Loss of p16INK4A expression was demonstrated in 55% of tumors (78 of 143) and was associated with reduced disease-free (P = 0.007) and overall (P = 0.014) survival. Multivariate analysis confirmed that in addition to pathological stage and regional lymph node status, cyclin D1 overexpression and loss of p16INK4A expression are independent predictors of death from tongue cancer. Loss of p16INK4A in the presence of cyclin D1 overexpression conferred a significantly worse disease-free (P = 0.011) and overall (P = 0.002) survival at 5 years. p53 nuclear accumulation and the Ki-67 labeling index were not prognostic. These data indicate that cyclin D1 overexpression and loss of p16INK4A expression predict early relapse and reduced survival in squamous cell carcinoma of the anterior tongue. Simultaneous assessment of cyclin D1 and p16INK4A protein levels define subgroups of patients at increased risk of relapse and may be of clinical utility in optimizing therapy.

Topics: Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Humans; Ki-67 Antigen; Lymphatic Metastasis; Male; Tongue Neoplasms; Tumor Suppressor Protein p53

Cyclin D1 in cooperation with its major catalytic partners, cyclin-dependent kinases cdk4 and cdk6, facilitates progression through the G1 phase of the eukaryotic cell cycle, in part through phosphorylation of the retinoblastoma protein. Cyclin D1's oncogenic properties have been suggested by its cooperation with ras or adenovirus E1a to transform cultured cells, as well its overexpression in transgenic mice that leads to breast cancer. Activated by a number of different mechanisms in human cancers, the cyclin D1 gene is frequently amplified in squamous epithelial cancers derived from the head/neck and esophageal regions. In order to study the functional consequences of cyclin D1 overexpression in these squamous epithelial specific sites, we have linked the Epstein-Barr virus ED-L2 promoter to the human cyclin D1 cDNA and utilized this transgene to generate founder lines. This transgene is transcribed specifically in the tongue, esophagus and forestomach, all sharing a stratified squamous epithelium. The transgene protein product localizes to the basal and suprabasal compartments of these squamous epithelial tissues, and mice from different lines develop dysplasia, a prominent precursor to carcinoma, by 16 months of age in contrast to age-matched wild-type mice. This transgenic model is useful in demonstrating cyclin D1 may be a tumor initiating event in aero-upper digestive squamous epithelial tissues.

Topics: Animals; Blotting, Southern; Cell Transformation, Neoplastic; Cyclin D1; Cyclins; DNA, Complementary; Esophageal Neoplasms; Esophagus; Gastric Mucosa; Herpesvirus 4, Human; Humans; Mice; Mice, Transgenic; Oncogene Proteins; Phenotype; Precancerous Conditions; Promoter Regions, Genetic; Stomach; Stomach Neoplasms; Tongue; Tongue Neoplasms