cyclin-d1 and Laryngeal-Neoplasms

Despite recent improvements in surgical and radiation therapy, failures still occur in patients with laryngeal squamous cell carcinomas, which may have a very different clinical outcome even when their clinical and histopathological characteristics are similar. The apparent inadequacy of "traditional" prognostic factors in predicting the clinical evolution of laryngeal squamous cell carcinomas has led to attempts to develop additional markers capable of distinguishing patients with a good prognosis from those who are more likely to relapse. A number of studies have demonstrated a relationship between tumourigenesis and alterations in the expression of cyclins, cyclin-dependent kinases and cyclin-dependent kinase inhibitors, but the data regarding laryngeal squamous cell carcinomas are somewhat conflicting. Herein a review is made of the published literature concerning the clinico-prognostic role of cyclin D1, D3 and p27, and personal data are described concerning laryngeal squamous cell carcinoma patients who underwent surgical resection at the ENT Department of the University of Milan. The results of our multivariate analyses demonstrated that cyclin D1, p27 and cyclin D3 overexpression are statistically significant predictors of disease-free survival (p = 0.0238, p = 0.0001 and p = 0.0217, respectively); the statistical correlation with overall survival was significant in the case of p27 (p = 0.0009) and cyclin D3 (p = 0.0189), and borderline in the case of cyclin D1 (p = 0.0622). In relation to cyclin D1/p27 coexpression, the patients with a cyclin D1-/p27+ phenotype showed the best prognosis, those with a cyclin D1/p27+ or cyclin D1-/p27- phenotype, an intermediate prognosis, and those with a cyclin D1+/p27- phenotype, the poorest prognosis (p = 0.0001 and p = 0.0001 for trend for disease-free survival; p = 0.0015 and p = 0.0008 for trend for overall survival). In the case of cyclin D1/cyclin D3 coexpression, the patients with cyclin D1+/cyclin D3+ tumours had the poorest overall survival, those with cyclin D1/cyclin D3+or cyclin D1+/cyclin D3- tumours showed intermediate course, and those with cyclin D1 /cyclin D3- tumours had the most favourable outcome (p = 0.0002). The findings of this review indicate that both types of cyclin D and p27 are involved in the genesis of laryngeal squamous cell carcinomas, and that immunohistochemical evaluations of biopsy samples may provide useful additional markers capable of identifying subgroups of patients with

Topics: Carcinoma, Squamous Cell; Cell Cycle Proteins; Cyclin D1; Cyclin D3; Cyclin-Dependent Kinase Inhibitor p27; Cyclins; Humans; Laryngeal Neoplasms; Tumor Suppressor Proteins

Some of the mechanisms involved in neoplastic transformation and progression of laryngeal squamous cell carcinoma (LSCC) are discussed. Although tumor suppressor inactivation of p53 and p16 is common in these tumors (about 50% each), oncogenic activation is less well characterized. Cyclin D1 and epidermal growth factor receptor amplification have been reported in one-third and one-quarter of LSCCs, respectively, both related to advanced stages, whereas c-myc could be amplified in 13% of cases although without associated overexpression. The role of ras in LSCC is, at most, exceptional, and the role of human papillomavirus infection in these neoplasms could have been largely overestimated. The AIS (amplified in squamous carcinoma) gene has been recently proposed as the main oncogenic target in head and neck squamous carcinomas and is a promising line of investigation. This, along with the link that exists between p53 and INK4 suppressor pathways through ARF and MDM-2, and the role of the universal cdk inhibitors (the Cip/Kip family) in these neoplasms deserve further investigation. Not forgotten are the mechanisms leading to cell immortalization and invasive capabilities acquisition, some of which are also briefly described.

Topics: Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Cyclin D1; Disease Progression; DNA, Neoplasm; Genes, p16; Genes, p53; Humans; Laryngeal Neoplasms; Molecular Biology; Proto-Oncogenes; Telomerase

As new knowledge of the molecular mechanisms of tumor development and progression is gained, studies are required to determine whether specific molecular genetic changes might be useful indicators in selecting patients for specific therapies. Alternatively, molecular or genetic changes may identify the patients at greatest risk for progression and relapse so that they can be treated with adjuvant therapy. p53 has an important function in the regulation of the cell cycle, DNA repair, and programmed cell death pathways. The Bcl-2 protein family has gained recent attention for its role in permitting or blocking apoptosis. Interaction between p53, Bcl-2, and other products of tumor suppressor genes or oncogenes are probably critical in tumor progression and response to treatment.

Topics: Apoptosis; Biomarkers, Tumor; Cell Cycle; Cyclin D1; Head and Neck Neoplasms; Humans; Laryngeal Neoplasms; Neoplasm Staging; Predictive Value of Tests; Prognosis; Proto-Oncogene Proteins c-bcl-2; Tumor Suppressor Protein p53

To determine the prognostic and predictive values of molecular marker expression profiles based on data from a randomized clinical trial of postoperative conventional fractionation (p-CF) therapy versus 7-day-per-week postoperative continuous accelerated irradiation (p-CAIR) therapy for squamous cell cancer of the head and neck.. Tumor samples from 148 patients (72 p-CF and 76 p-CAIR patients) were available for molecular studies. Immunohistochemistry was used to assess levels of EGFR, nm23, Ki-67, p-53, and cyclin D1 expression. To evaluate the effect of fractionation relative to the expression profiles, data for locoregional tumor control (LRC) were analyzed using the Cox proportional hazard regression model. Survival curves were compared using the Cox f test.. Patients who had tumors with low Ki-67, low p-53, and high EGFR expression levels and oral cavity/oropharyngeal primary cancer sites tended to benefit from p-CAIR. A joint score for the gain in LRC from p-CAIR based of these features was used to separate the patients into two groups: those who benefited significantly from p-CAIR with respect to LRC (n = 49 patients; 5-year LRC of 28% vs. 68%; p = 0.01) and those who did not benefit from p-CAIR (n = 99 patients; 5-year LRC of 72% vs. 66%; p = 0.38). The nm23 expression level appeared useful as a prognostic factor but not as a predictor of fractionation effect.. These results support the studies that demonstrate the potential of molecular profiles to predict the benefit from accelerated radiotherapy. The molecular profile that favored accelerated treatment (low Ki-67, low p-53, and high EGFR expression) was in a good accordance with results provided by other investigators. Combining individual predictors in a joint score may improve their predictive potential.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; ErbB Receptors; Female; Head and Neck Neoplasms; Humans; Ki-67 Antigen; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Neoplasm Recurrence, Local; NM23 Nucleoside Diphosphate Kinases; Oropharyngeal Neoplasms; Proportional Hazards Models; Radiotherapy; Radiotherapy Dosage; Risk Factors; Tumor Suppressor Protein p53

In a previous trial, we found that combined 13-cis-retinoic acid, IFN-alpha, and alpha-tocopherol more effectively reversed advanced premalignant lesions of the larynx than of the oral cavity and that cyclin D1 (CD1) G/A870 single nucleotide polymorphism correlated with cancer risk. We conducted the present trial primarily to confirm the clinical activity of the combination in advanced laryngeal premalignancy and to confirm and extend our findings on CD1, both genotype and protein expression, in association with cancer risk in this setting. Twenty-seven moderate-to-severe laryngeal dysplasia patients underwent induction with combined 13-cis-retinoic acid daily, alpha-IFN twice weekly, and alpha-tocopherol daily for 1 year; 14 nonprogressing patients then were randomized to maintenance fenretinide or placebo for 2 years. During induction, two patients had pathologic complete responses, six had partial responses (30% overall response rate), and five developed laryngeal cancer. There were no significant differences between maintenance fenretinide and placebo in response or cancer rates. Ten patients developed cancer overall. Twenty-four patients were evaluated for the CD1 G/A870 genotype, and 23 for pretreatment and posttreatment CD1 protein expression. Consistent with our earlier report, shorter cancer-free survival was associated with the CD1 AA/AG genotype (P = 0.05). Extending our earlier work, high CD1 expression was associated with worse cancer-free survival overall (P = 0.04) and within each CD1 genotype group. These findings support CD1 genotype and protein expression as important risk markers for laryngeal cancer and suggest future trials targeting upstream regulators of CD1 transcription.

Topics: alpha-Tocopherol; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Cyclin D1; Female; Fenretinide; Gene Expression; Genotype; Humans; Immunohistochemistry; Interferon-alpha; Isotretinoin; Laryngeal Diseases; Laryngeal Neoplasms; Male; Polymorphism, Single Nucleotide; Precancerous Conditions

Altered cyclin D1 expression in advanced preinvasive lesions of the upper aerodigestive tract (UADT) is associated with an increased risk of developing cancer and histologic progression during and after combination biochemopreventive therapy (13-cis-retinoic acid, alpha-interferon, and alpha-tocopherol). Both alleles of the adenine (A)/guanine (G) cyclin D1 polymorphism located at nucleotide 870 encode two alternatively spliced transcripts, but the A allele preferentially encodes a protein with an extended half-life. We investigated whether the cyclin D1 genotype at nucleotide 870 was associated with baseline levels of cyclin D1 protein, post-treatment modulation of cyclin D1 protein levels, histologic response to treatment, and the outcome for subjects with preinvasive UADT lesions after biochemopreventive therapy.. UADT tissue biopsy samples were obtained before and 6 and 12 months after biochemopreventive treatment from 31 individuals with advanced preinvasive UADT lesions. Tissues were examined for cyclin D1 genotype (by DNA single-strand conformation polymorphism analysis), for cyclin D1 protein expression (by immunohistochemistry), and for cyclin D1 gene copy number (by fluorescence in situ hybridization). Associations of cyclin D1 genotype with histologic response to therapy and time to progression to a higher degree of dysplasia or invasive cancer were investigated. All statistical tests were two-sided.. The A allele was associated with increased baseline cyclin D1 expression in the parabasal epithelial layer (16 of 18 AA/AG subjects versus four of nine GG subjects; P =.02), decreased histologic response to biochemopreventive treatment (six of 21 AA/AG subjects versus four of 10 GG subjects; P =.70), decreased favorable modulation of cyclin D1 expression by the treatment (seven of 18 AA/AG subjects versus eight of nine GG subjects; P =.02), and shorter progression-free survival (P =.05).. The cyclin D1 A allele was associated with a diminished modulation of normal physiologic and treatment-induced decreased expression of cyclin D1, a decreased likelihood of response to biochemopreventive intervention, and an increased rate of progression to cancer development, findings that require validation in a larger cohort.

Topics: alpha-Tocopherol; Antineoplastic Combined Chemotherapy Protocols; Cyclin D1; Disease Progression; Gene Expression Regulation, Neoplastic; Genotype; Humans; Immunohistochemistry; Immunologic Factors; In Situ Hybridization, Fluorescence; Interferon-alpha; Isotretinoin; Laryngeal Neoplasms; Mouth Neoplasms; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Prospective Studies; Treatment Outcome

Cyclin D1 is a set of periodic protein which governs G1 progression. Cyclin D1 gene is localized on chromosome 11q13 which encodes a 295-aa protein. Overexpression of cyclin D1 leads to abnormal cellular proliferation. Which underlies processes of tumorigenesis. In this paper twenty-five fresh specimens of laryngeal carcinomas were examined by means of immune flurescence technique. Overexpression of cyclin D1 was found in 9 of 16 laryngeal carcinomas (37%). There was no statistical correlation between overexpression of cyclin D1 and TNM staging, differentiation grading (P > 0.05). Normal tissue adjacent to tumors lacked any detectable cyclin D1 expression or rare scattered positive cells. It was likely that cyclin D1 overexpression wasn't an early event in processes of tumorigenesis and tumor progression. Follow-up investigation demonstrated that tumors recurred in 4 of 9 primary tumors overexpressing cyclin D1. But only 1 of 16 that expressed cyclin D1-negative. There was statistical significance between them, so overexpression of cyclin D1 could serve as a new prognostic marker.

Topics: Cyclin D1; Double-Blind Method; Follow-Up Studies; Gene Expression; Genes, bcl-1; Humans; Laryngeal Neoplasms; Neoplasm Recurrence, Local; Prognosis

As an important member of Wnt/β-catenin signaling pathway, the aberrant expression of β-catenin has been implicated in many cancers. Chibby, a β-catenin binding partner, is an antagonist involved in this pathway. In contrast, thyroid cancer 1 (TC1) as an activator of this pathway can relieve the antagonistic activity of Chibby on the β-catenin-mediated transcription and is high expressed in human tumors. The objectives of this study were to examine the expression of TC1, Chibby, and β-catenin and investigate the association among them in laryngeal squamous cell carcinoma (LSCC). The expression of TC1, Chibby, β-catenin, c-Myc, Cyclin D1, and matrix metalloproteinase-7 (MMP-7) were examined by immunohistochemistry in samples from 53 LSCC patients. Compared with normal laryngeal squamous epithelium (NLSE), there were upregulated expression of TC1, downregulated expression of Chibby, and aberrant cytoplasmic expression of β-catenin in the LSCC tissues (P < .001). The high expression of TC1 was correlated with the tumor site, advanced TNM and T stage, lymphovascular invasion, and poor differentiation in LSCC tissues (P < .050). There were correlations between the aberrant expression of β-catenin and the tumor site, advanced TNM and T stage, lymphovascular invasion, perineurial invasion, and poor differentiation in LSCC tissues (P < .050). Upregulated TC1 and downregulated Chibby were correlated with aberrant expression of β-catenin (P < .001), but no correlation between them (P = .076). The percent of abnormal expression of β-catenin in LSCC was 96.00% in TC1+/Chibby-, 73.68% in TC1+/Chibby+, 0.00% in TC1-/Chibby-, and 0.00% in TC1-/Chibby + group (P < .001). High expression of c-Myc, Cyclin D1, and MMP-7 was observed in LSCC tissues (P < .001). There was statistically significant about the expression of Cyclin D1 and MMP-7 among the groups of TC1+/Chibby-, TC1+/Chibby+, TC1-/Chibby-, and TC1-/Chibby + (P < .001), but was not significance about the expression of c-Myc among them (P = .339). No association was found between overall survival and the expression of TC1, Chibby, and β-catenin (P > .05). The upregulated expression of TC1 and downregulated expression of Chibby were correlated with the aberrant expression of β-catenin and the high expression of Cyclin D1 and MMP-7 in LSCC tissues.

Topics: beta Catenin; Carrier Proteins; Cyclin D1; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Laryngeal Neoplasms; Matrix Metalloproteinase 7; Nuclear Proteins; Squamous Cell Carcinoma of Head and Neck

This study investigated the role of lncRNA CASC15 in laryngeal squamous cell carcinoma (LSCC).. This study included 58 LSCC patients. Both tumor (LSCC) and adjacent (within 3 cm around tumors) non-tumor tissues from 3 different sites of each patient were collected. CCK-8 assay was used to determine cell proliferation. The expression levels of proteins and mRNAs were determined by Western blotting analysis and qRT-PCRs, respectively.. CASC15 was upregulated in LSCC and high expression levels of CASC15 predicted poor survival. In LSCC tissues, CASC15 was negatively correlated with miR-365 but positively correlated with cyclin D1. In LSCC cells, overexpression of CASC15 resulted in downregulation of miR-365 and upregulation of cyclin D1. Overexpression of miR-365 did not affect the expression of CASC15 but downregulated cyclin D1. Overexpression of Cyclin D1 did not affect the expression of miR-365 and CASC15. Overexpression of CASC15 and cyclin D1 led to promoted, while overexpression of miR-365 led to inhibited LSCC cell proliferation. In addition, overexpression of miR-365 reduced the effects of overexpression of CASC15.. Therefore, CASC15 upregulates cyclin D1 by downregulating miR-365 in LSCC to promote cell proliferation.

Topics: Carcinoma, Squamous Cell; Cell Proliferation; Cyclin D1; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Laryngeal Neoplasms; MicroRNAs; RNA, Long Noncoding; Squamous Cell Carcinoma of Head and Neck; Up-Regulation

The membrane EGFR (mEGFR) protein overexpression in the head and neck squamous cell carcinoma (SCC) is considered to cause increased EGFR activity which adds to tumorigenicity and therapy resistance. The mEGFR upon stimulation can translocate to the nucleus nuclear EGFR (nEGFR) where it has been associated with poor prognosis and worse survival in many cancers. The relevance of differentially located EGFR proteins in laryngeal lesions has not been studied enough and remains unclear. Aim of our study was to examine nEGFR and mEGFR protein expression as well as EGFR gene status and cell cycle proliferation markers in the laryngeal polyps, dysplasia, and SCC using immunohistochemistry and in situ hybridization. There was significantly higher frequency of strong nEGFR between SCC, dysplasia, and polyps (P<0.0001), and strong mEGFR in the SCC and laryngeal dysplasia comparing to polyps (P<0.0001). Gene amplification was confirmed only in relatively small number of SCC but not in non-neoplastic lesions. In dysplasia the statistically significant positive correlations between nEGFR, and Ki-67 (P=0.029), p53 (P=0.001), and cyclin D1 (P=0.031) were found. nEGFR and mEGFR expression showed statistically significant inverse correlation in the SCC (P=0.004) as well as nEGFR and cyclin D1 (P=0.032). Univariate statistical analysis showed statistically significant correlation between strong nEGFR protein expression and worse overall survival in laryngeal SCC, alone or in coexpression with strong cyclin D1 and high Ki-67 (P=0.025, P=0.046, P=0.043, respectively). Our data show that nEGFR cellular localization might influence biology of the laryngeal carcinogenesis and is indicator of poor survival.

Topics: Aged; Cell Nucleus; Cyclin D1; Disease-Free Survival; ErbB Receptors; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Survival Rate

Cyclin D1 (CCND1) is a well-known proliferation promoter that accelerates G1/S transition in cancer. However, the underlying mechanism by which CCND1 is regulated is still largely unknown. In this study, we identified a novel circular RNA (circRNA) derived from CCND1 (circ-CCND1, hsa_circ_0023303) as a key regulator for CCND1. circ-CCND1 was found to be markedly up-regulated in laryngeal squamous cell carcinoma (LSCC) and closely associated with aggressive clinical features and adverse prognosis. Depletion of circ-CCND1 significantly inhibited LSCC cell proliferation in vitro and retarded tumour growth in vivo. Regarding the mechanism, circ-CCND1 physically bound to human antigen R (HuR) protein to enhance CCND1 mRNA stability; on the other hand, circ-CCND1 could act as an effective sponge for miR-646 to alleviate the repression of miR-646 on CCND1 mRNA. As a result, circ-CCND1 post-transcriptionally elevated CCND1 expression via coordinated avoidance of CCND1 mRNA decay, thereby promoting LSCC tumorigenesis. Taken together, our findings uncover the essential proliferation-promoting role of circ-CCND1 through regulation of the stability of CCND1 mRNA in LSCC. Targeting circ-CCND1 may be a promising treatment for LSCC patients.

Topics: Carcinogenesis; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cyclin D1; ELAV-Like Protein 1; Female; Gene Expression Regulation, Neoplastic; Humans; Laryngeal Neoplasms; Male; MicroRNAs; Middle Aged; Prognosis; RNA, Circular; RNA, Messenger; Up-Regulation

Deregulation of critical proteins involved in cell cycle stability, such as cyclins, is a frequent genetic event in the development and progression of solid malignancies. Concerning laryngeal squamous cell carcinoma (LSCC), cyclin D1 oncogenic transformation leads to an aberrant protein expression and seems to affect the biological behaviour of the neoplasm. The aim of this study was to determine the correlation of cyclin D1 numerical imbalances with the corresponding protein expression levels in LSCCs.. Using tissue microarray (TMA) technology, fifty (n=50) histologically confirmed primary LSSCs were cored at a diameter of 1.5 mm. Immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) analyses were applied. Concerning the screening process in CISH slides, a novel real-time reference and calibration grid platform was implemented.. Protein overexpression was observed in 22/50 (44%) cases; whereas, gene amplification was seen in 13/50 (26%) cases (p=0.02). Combined protein/ gene deregulation was associated with the stage of malignancy (p= 0.0014, p=0.001), whereas overall protein expression was strongly correlated with the grade of tumour (p= 0.001).. Cyclin D1 gene amplification led to aberrant protein expression in LSCCs and it was also correlated with an aggressive biological behaviour. To best of our knowledge, this study was the first described grid based CISH analysis under conventional bright field microscopy for detecting gene numerical imbalances while providing a novel and accurate description for screening-mapping process in the entire slide area.
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Topics: Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Female; Follow-Up Studies; Gene Amplification; Humans; Laryngeal Neoplasms; Male; Middle Aged; Prognosis; Tissue Array Analysis

Laryngeal cancer, one of the most common head and neck malignancies, is an aggressive neoplasm. Increasing evidence has demonstrated that microRNAs (miRNAs) exert important roles in oncogenesis and progression of diverse types of human cancers. miR-632, a tumor-related miRNA, has been reported to be dysregulated and implicated in human malignancies; however, its biological role in laryngeal carcinoma remains to be elucidated. The present study aimed at exploring the role of miR-632 in laryngeal cancer and clarifying the potential molecular mechanisms involved. In the current study, miR-632 was found to be significantly upregulated both in laryngeal cancer tissues and laryngeal cancer cell lines. Functional studies demonstrated that miR-632 accelerated cell proliferation and colony formation, facilitated cell migration and invasion, and enhanced the expression of cell proliferation-associated proteins, cyclin D1 and c-myc. Notably, miR-632 could directly bind to the 3'-untranslated region (3'-UTR) of glycogen synthase kinase 3β (GSK3β) to suppress its expression in laryngeal cancer cells. Mechanical studies revealed that miR-632 promoted laryngeal cancer cell proliferation, migration, and invasion through negative modulation of GSK3β. Pearson's correlation analysis revealed that miR-632 expression was inversely correlated with GSK3β mRNA expression in laryngeal cancer tissues. Taken together, our findings suggest that miR-632 functions as an oncogene in laryngeal cancer and may be used as a novel therapeutic target for laryngeal cancer.

Topics: 3' Untranslated Regions; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Down-Regulation; Glycogen Synthase Kinase 3 beta; Humans; Laryngeal Neoplasms; MicroRNAs; Neoplasm Invasiveness; Proto-Oncogene Proteins c-myc; Real-Time Polymerase Chain Reaction; RNA, Neoplasm; Up-Regulation

Topics: Aged; Base Pairing; Base Sequence; Carcinogens; Carcinoma, Squamous Cell; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Disease Progression; Epithelial Cells; Female; Gene Expression Regulation, Neoplastic; Humans; Laryngeal Neoplasms; Male; MicroRNAs; Middle Aged; Neoplasm Staging; RNA, Circular; RNA, Small Interfering; Signal Transduction

To evaluate CD163+ tumor-associated macrophages (TAMs), Ki-67, and cyclin D1 to differentiate laryngeal dysplasia in the 2017 World Health Organization classification.. Immunohistochemistry for CD163, Ki-67, and cyclin D1 was performed using paraffin-embedded specimens. CD163+ TAMs infiltrating the epithelium were estimated. Ki-67 and cyclin D1 were evaluated in four parts of the epithelium-basal, parabasal, middle third, and upper third layers.. In total, 133 specimens were analyzed, including low-grade dysplasia (n = 31), high-grade dysplasia (n = 49), carcinoma in situ (n = 23), and normal mucosa (n = 30). CD163+ TAMs infiltrating the epithelium were significantly higher in high-grade dysplasia than in low-grade dysplasia. In the basal layer, Ki-67+ and cyclin D1+ cells were overexpressed in high-grade dysplasia (P < .0001). The area under the curve was 0.958 for Ki-67 and 0.909 for CD163+ TAMs (P < .0001).. CD163+ TAMs infiltrating the epithelium and Ki-67 overexpression in the basal layer may serve as biomarkers to differentiate low-grade dysplasia from high-grade dysplasia of the larynx. A symmetric proliferative pattern was observed during laryngeal carcinogenesis following Ki-67 overexpression.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biomarkers, Tumor; Carcinoma in Situ; Cyclin D1; Humans; Immunohistochemistry; Ki-67 Antigen; Laryngeal Diseases; Laryngeal Mucosa; Laryngeal Neoplasms; Macrophages; Receptors, Cell Surface

Laryngeal carcinoma is one of the most common head and neck cancers. MicroRNAs are a class of small non-coding RNAs 18-25 nucleotides in length that post-transcriptionally regulate gene expression and have a pivotal role in many biological processes including cancer development. In this study, we investigated the role of miR-34a in laryngeal carcinoma and confirmed the regulation of cyclin D1 (CCND1) by miR-34a. We examined miR-34a expression levels in 71 laryngeal carcinoma patient specimens by quantitative reverse transcription-PCR, and analyzed the clinicopathological significance of the obtained data. Then, functional assays were performed to investigate the potential effects of miR-34a on cancer cell proliferation and migration. In addition, western blotting, luciferase reporter assay and several algorithms were conducted to confirm that CCND1 is directly regulated by miR-34a. We demonstrated that the miR-34a expression level was significantly downregulated in laryngeal carcinoma clinical specimens compared with that observed in their paired adjacent normal tissues. Additionally, miR-34a expression was also inversely correlated with lymph node metastasis and clinical stage. Functional assays showed that ectopic expression of miR-34a inhibited cell proliferation and migration in laryngeal carcinoma cells. Bioinformatic analysis identified CCND1 as a potential target of miR-34a. Moreover, we confirmed that miR-34a inhibited the expression of CCND1 by directly binding to its 3'-untranslated region. Silencing of CCND1 induced effects similar to those of miR-34a ectopic expression, and in laryngeal carcinoma tissues, miR-34a and CCND1 were inversely correlated. Our data suggest that tumor suppressor miR-34a could serve as a new potential diagnostic marker and that ectopic expression of miR-34a may be used as a therapeutic target for laryngeal carcinoma.

Topics: 3' Untranslated Regions; Biomarkers, Tumor; Carcinoma; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Down-Regulation; Female; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Humans; Laryngeal Neoplasms; Lymphatic Metastasis; Male; MicroRNAs; Middle Aged

To investigate the effects of a dual phosphoinosmde-3-kinase (PI3K)/ mammalian target of rapamycin (mTOR) inhibitor, PI-103, cooperating with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the laryngeal squamous carcinoma Hep-2 cells.. Hep-2 cells were divided into 7 groups: LY294002 group, Rapamycin group, PI-103 group, LY294002+ TRAIL group, Rapamycin+ TRAIL group, PI-103+ TRAIL group and control group.The cell cycle and apoptosis of Hep-2 cells were assessed by flow cytometry.For PI-103 group, PI-103+ TRAIL group and control group, migration and invasion ability were measured by transwell migration and invasion assay respectively.The expression of relative proteins in apoptosis and PI3K/AKT/mTOR signal pathway was examined by Western blotting.. Combination of PI-103 and TRAIL could make cell cycle arrest at S phase (G1: 1.80%±0.30%; G2: 0.00), inhibit cell proliferation, and enhance apoptosis (66.78%±2.93%) (P<0.05). Combination of PI-103 and TRAIL could statistically decrease the migration and invasion number of Hep-2 cells (17.0±3.4, 18.4±5.4) than that of PI-103 group (41.2±3.8, 41.6±4.7). PI-103 could inhibit PI3K/AKT/mTOR signal pathway by decreasing the protein expression of p-AKT and p-4E-BP1.Comparing with the control group, the expression of cysteinyl aspartate specific proteinase (Caspase) 9, 8, 3 were increased while the expression of Cyclin D1, Cyclin E1, p-AKT, p-4E-BP1 were decreased in PI-103 and PI-103+ TRAIL group (P<0.05).. Enhanced anti-tumor effects was observed by combination of PI-103 and TRAIL on laryngeal cancer cells in vitro and this combined administration might be a promising strategy for clinical treatment of laryngeal cancer.

Topics: Animals; Apoptosis; Carcinoma, Squamous Cell; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Chromones; Cyclin D1; Cyclin E; Furans; Head and Neck Neoplasms; Humans; Laryngeal Neoplasms; Morpholines; Oncogene Proteins; Phosphatidylinositol 3-Kinases; Pyridines; Pyrimidines; Signal Transduction; Squamous Cell Carcinoma of Head and Neck; TNF-Related Apoptosis-Inducing Ligand; TOR Serine-Threonine Kinases

The cell surface glycoprotein Trop2 is overexpressed in various types of epithelial cancer. Laryngeal carcinoma is one of the most common types of head and neck cancer and in a previous study, the expression of Trop2 in laryngeal squamous cell carcinoma (LSCC) was identified as an independent prognostic factor. However, the biological significance of Trop2 in LSCC development remains unclear. In the current study, Trop2 protein expression in fresh LSCC tissue and paracancerous tissue was investigated using western blotting. Trop2 in the Hep2 laryngeal cell line was subsequently suppressed by transfection with small interfering RNA (siRNA). The effects of knockdown of Trop2 on cell viability, migration, invasiveness and ERK/MAPK pathway activity were investigated in the current study. The expression of Trop2 in fresh LSCC tissue was demonstrated to be significantly greater than that in paracancerous tissue. Trop2 expression was also identified to be required for proliferation, migration and invasiveness of Hep2 laryngeal carcinoma cells, as all were blocked by siRNA-mediated Trop2 inhibition. Notably, the ERK/MAPK signaling pathway and cell cycle factor, cyclin D1, were identified to be suppressed following the knockdown of Trop2 in Hep2 cells. These observations suggest that Trop2 serves an oncogenic role in LSCC and has potential as a therapeutic target.

Topics: Antigens, Neoplasm; Carcinoma, Squamous Cell; Cell Adhesion Molecules; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cyclin D1; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Laryngeal Neoplasms; Mitogen-Activated Protein Kinase 1; Neoplasm Invasiveness; Signal Transduction

To observe the effect of metastasis-associated gene 1 (MTA1) on the growth and metastasis of laryngeal squamous cell carcinoma in nude mice using RNA interference technology (RNAi).. Lentiviral vector of short hairpin RNA (shRNA) targeting MTA1 was constructed and packaged to transfect Hep-2 cells. Hep-2 cells transfected with scramble shRNA and MTA1 shRNA were injected into the paw pad of nude mice (n=5 per group). Nine weeks after modeling of the lymphatic metastasis of laryngeal carcinoma, the mice were sacrificed, and tumor tissues and inguinal lymph node were harvested to be subjected to HE staining, reverse transcription PCR and Western blotting.. The gene screening showed the lentiviral vector of MTA1 shRNA was constructed successfully, and those tumor cells were transplanted and grew well in all mice. The size of tumor in the mice of MTA1 shRNA tansfected group was obviously smaller compared to the scramble shRNA transfected group at the same time point. No inguinal lymph node metastasis was found in the mice of MTA1 shRNA group. In contrast, the tumor cells were seen in the inguinal lymph nodes of the scramble shRNA infected mice. Reverse transcription PCR and Western blotting showed that the mRNA and protein levels of MTA1, β-catenin, matrix metalloproteinase-9 (MMP-9), cyclin D1 were obviously reduced in MTA1 shRNA infected mice compared with the scramble shRNA infected mice.. The inhibition of MTA1 gene could depress the growth and metastasis of laryngeal squamous cell carcinoma in nude mice.

Topics: Animals; beta Catenin; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Female; Head and Neck Neoplasms; Laryngeal Neoplasms; Lymphatic Metastasis; Matrix Metalloproteinase 9; Mice; Mice, Nude; Repressor Proteins; RNA, Small Interfering; Squamous Cell Carcinoma of Head and Neck; Trans-Activators; Transcription Factors

The aberrant expression of microRNAs (miRNAs) has been found in various types of cancer. miR-205 was reported to be upregulated in laryngeal squamous cell carcinoma (LSCC) tissues, however, the mechanisms by which miR-205 functions as a regulator of LSCC are largely unknown.. In this study, Real-time qPCR and Western blot assay showed that expression of miR-205 was upregulated and expression of cyclin-dependent kinase 2-associated protein 1 (CDK2AP1) was downregulated in LSCC tissues. The expression levels of miR-205 were negatively related to those of CDK2AP1 in LSCC tissues and cell lines. Moreover, we found that miR-205 was the upstream regulator of CDK2AP1 and could suppress the CDK2AP1 expression in LSCC cells. 3-(4,5-dimethylthiazal-2-yl)-2,5-diphenyl-tetrazolium bromide assays and transwell invasion assay were performed to test the proliferation and invasion of LSCC cells. Gelatin zymography was used to detect the activity of MMP2 and MMP9. CDK2AP1, c-Myc and CyclinD1 expression in cells was assessed with Western blotting. We found that miR-205 was the upstream regulator of CDK2AP1 and could suppress the expression of CDK2AP1 in LSCC cells. In addition, miR-205 significantly induced cell proliferation and invasion by suppressing CDK2AP1 expression. Consistent with miR-205 inhibitors, overexpressed CDK2AP1 suppressed the activity of MMP2 and MMP9 and c-Myc and CyclinD1 expression in LSCC cells.. These findings help us to better elucidate the molecular mechanisms of LSCC progression and provide a new theoretical basis to further investigate miR-205 as a potential biomarker and a promising approach for LSCC treatment.

Topics: Biomarkers, Tumor; Blotting, Western; Carcinoma, Squamous Cell; Cell Proliferation; Cyclin D1; Down-Regulation; Gene Expression Regulation, Neoplastic; Genes, myc; Hep G2 Cells; Humans; Laryngeal Neoplasms; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; MicroRNAs; Neoplasm Invasiveness; Primary Cell Culture; Real-Time Polymerase Chain Reaction; Suppression, Genetic; Tumor Suppressor Proteins

Head and neck squamous cell carcinomas (HNSCCs) are characterized by marked heterogeneity in their biological behavior and response to treatment. Our goal was the identification of biomarkers that can be used to predict response to chemotherapy in these patients.. The expression of EGFR, p53, Cyclin D1, p16, p21, p27, p-AKT, HIF-1α, Caspase 3 and BCL2 was analyzed by immunohistochemistry in 41 primary laryngeal/hypopharyngeal squamous cell carcinomas of patients that received induction chemotherapy (cisplatin and 5-fluorouracil) as part of their treatment.. Positive expression of p27 and BCL2 had a significant predictive value for chemotherapy response in univariate analysis. The combination of both proteins was not superior in predicting the response to chemotherapy. Furthermore, p27 expression was the only significant predictor of chemotherapy response in multivariate analysis (P=0.015).. p27 Expression may serve as predictive biomarker of response to induction chemotherapy in HNSCC patients.

Topics: Adult; Aged; Antimetabolites, Antineoplastic; Antineoplastic Agents; Biomarkers, Tumor; Carcinoma, Squamous Cell; Caspase 3; Cisplatin; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; ErbB Receptors; Female; Fluorouracil; Humans; Hypopharyngeal Neoplasms; Hypoxia-Inducible Factor 1, alpha Subunit; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Proteins; Prognosis; Proliferating Cell Nuclear Antigen; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Tumor Suppressor Protein p53

Cyclin D1 and E-cadherin are important factors in the progression and metastasis of cancers. Their role in laryngeal carcinoma has been studied with conflicting results. To define the frequency of cyclin D1 and E-cadherin expression and its correlation with both the clinicopathological characteristics and prognosis of patients with laryngeal squamous cell carcinoma (LSCC). Tumor tissue samples from 75 patients with laryngeal squamous cell carcinoma were examined for cyclin D1 and E-cadherin expression by immunohistochemistry. The relationship between the expression of both molecules and the age and sex of the patient, tumor site, tumor differentiation, lymph node metastasis, tumor invasiveness, TNM stages, tumor recurrence and overall survival was analyzed. Cyclin D1 was found to be a significant independent prognostic factor of lymph node metastasis (p = 0.000). The multivariate analysis revealed that cyclin D1 and E-cadherin expression wasn't an independent prognostic factor of local recurrence free survival (LRFS) in patients with LSCC (P = 0.56 and 0.28) respectively. However, the univariate analysis revealed a significant association between them and LRFS (p = 0.003 and 0.000) respectively. Also, the group of high cyclin D1 /low E-cadherin expression had the poorest prognosis, so they might serve as potential predictors of the prognosis of the patients with LSCC. E-cadherin was found to affect the overall survival (OAS) significantly by the univariate analysis (p = 0.01). However, by the multivariate analysis the TNM stage was the only independent prognostic factor of OAS (p < 0.05). Cyclin D1 can be used as an independent prognostic marker of lymph node metastasis in patients with LSCC and can help to identify those patients with clinically negative lymph nodes but with considerable risk for occult metastasis. Detection of cyclin D1 and E-cadherin status in LSCC may contribute to the identification of patients with high risk factors of local recurrence. However, they don't appear to be better prognostic predictors than other established markers in LSCC.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cadherins; Carcinoma, Squamous Cell; Cyclin D1; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Grading; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Survival Rate

The histopathology of premalignant laryngeal lesions does not provide reliable information on the risk of malignant transformation, hence we examined new molecular markers which can easily be implemented in clinical practice. Dual-target fluorescence in situ hybridisation (FISH) for chromosome 1 and 7 centromeres was performed on tissue sections of laryngeal premalignancies in 69 patients. Chromosome instability was indicated by numerical imbalances and/or polysomy for chromosomes 1 and 7. Additionally, immunostainings for p53, Cyclin D1 and (p)FADD expression were evaluated. Malignant progression was recorded. Eighteen patients with carcinoma in situ (CIS) were treated after diagnosis and excluded from follow-up. Chromosome instability was strongly associated with a high risk of malignant transformation, especially in lower grade lesions (hyperplasia, mild and moderate dysplasia; odds ratio = 8.4, p = 0.004). Patients with lesions containing chromosome instability showed a significantly worse 5-year progression-free survival than those with premalignancies without chromosome instability (p = 0.002). Neither histopathology nor the protein markers predicted progression in univariate analysis, although histopathological diagnosis, p53 and FADD contributed positively to chromosome instability in multivariate analysis. Chromosome instability is associated with malignant progression of laryngeal premalignancies, especially in lower grade lesions. These results may contribute to better risk counselling, provided that they can be validated in a larger patient set.

Topics: Adult; Aged; Chromosomal Instability; Cyclin D1; Disease Progression; Disease-Free Survival; Fas-Associated Death Domain Protein; Female; Humans; Hyperplasia; In Situ Hybridization, Fluorescence; Kaplan-Meier Estimate; Laryngeal Neoplasms; Larynx; Male; Middle Aged; Precancerous Conditions; Young Adult

Chibby (Cby) inhibits Wnt/β-catenin-mediated transcriptional activation by competing with Lef-1 (the transcription factor and target of β-catenin) to bind to β-catenin. This suggests that Cby could be a tumor suppressor protein. In the present study, we examined Cby expression in laryngeal squamous cell carcinoma (LSCC) and its function and mechanism in laryngeal carcinoma cell lines. Cby expression levels were investigated by immunohistochemistry in a panel of 36 LSCC patient cases. The expression of β-catenin, c-myc and cyclin D1 in Hep-2 were determined through RT-PCR and western blot analysis. Activity of Wnt/β-catenin signaling pathway after overexpression of Cby was measured by TCF/LEF luciferase reporter gene assay. Proliferation, clone forming ability, cell cycle distribution and cell apoptosis of Hep-2 cells were detected by MTT assay, plate colony forming assay, flow cytometry and TUNEL assay, respectively. This study showed that expression of Cby protein was strongly downregulated in LSCC tumor tissues in comparison to normal laryngeal mucosa samples. No significant correlation was found between the expression of Cby in tumor tissue and gender, age, clinical stage and tumor differentiation of laryngeal cancer patients. When Cby was overexpressed in Hep-2 cells, the expression of cyclin D1 was reduced and β-catenin activity was inhibited. Proliferation and plate colony forming assays revealed a significant inhibitory effect of Cby on growth and colony formation ability of Hep-2 cells after Cby overexpression in comparison to control and mock-infected cells. In addition, we also found that upregulated expression of Cby resulted in accumulation of numbers of cells in G0/G1 phase with concomitant decrease in S phase by cell cycle assay. TUNEL staining demonstrated that, compared with the control group, the rate of apoptosis in the plv-cs2.0-Cby group was significantly increased. Taken together, downregulation of Cby was observed in LSCC, but with no significant correlation to the clinicopathological features of LSCC patients. Overexpression of Cby effectively suppressed laryngeal carcinoma cell growth and promoted its apoptosis. A better understanding of the mechanisms of Cby gene activation in LSCC could provide potential novel therapeutic targets for human laryngeal carcinoma.

Topics: Adult; Aged; Apoptosis; Carcinoma, Squamous Cell; Carrier Proteins; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Hep G2 Cells; Humans; Laryngeal Neoplasms; Male; Middle Aged; Nuclear Proteins; Wnt Signaling Pathway

To investigate the expression and clinicopathological significance of the cell cycle regulators cyclin E, cyclin D1, p21, p16 in laryngeal carcinogenesis tissus.. The expression of cell cycle regulators were detected by flow cytometry method in 23 cases of polyps of vocal cord, 69 cases of laryngeal precancerous change and 33 cases of laryngeal squamous cell carcinoma (LSCC), which tissue was paraffin embedded, sliced, dewaxed, and prepared into the cell suspension, then fluorescently labeled by cyclin E, cyclin D1, p21 and p16.. In polyps of vocal cord, laryngeal precancerous change and LSCC, The positive expression rate of cyclin E and cyclin D1 were respectively 13.04%, 20.29D, 42.420 and 26.09%, 43.48% and 93.94%. The positive expression rate of p16 and p21 were respectively 61.90%, 40.98%, 14.28% and 47.62%, 23.81%, 26.23%. Those showed the positive expression rate of cyclin D1, cyclin E gradually decreased from vocal cord polyps, laryngeal precancerous change to LSCC, (P < 0.05, P < 0.01), while the positive expression rate of p21 and p16 gradually decreased (P < 0.01).. The abnormal expression of cell cycle regulatory factors is the molecular events of laryngeal carcinoma. High expression of positive regulatory factors cyclin D1 and cyclin E, and low expression of negative regulatory factors p16 and p21, which showed the imbalance of multiple positive and negative regulatory factors related with cell cycle play an important role in the occurrence of laryngeal cancer.

Topics: Adult; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; Female; Flow Cytometry; Humans; Laryngeal Neoplasms; Male; Middle Aged; Oncogene Proteins

Analyzing chromosomal amplifications delivers valuable information for identification of oncogenes. For carcinomas of the oral cavity only few genes have been identified in amplified regions. The aim of this study was to search genes in amplified regions as possible biomarkers and targets for novel therapies.. DNA from 10 carcinomas of the floor of the oral cavity was examined using a 500K Array GeneChip (Affymetrix 6.0) to detect chromosomal losses, gains or amplifications. Suspicious alterations were validated on tissue microarrays using fluorescence in situ hybridization (FISH) with respective probes.. FISH-validation on tissue arrays confirmed PPFIA1 amplifications as one of the most frequent events (32.6%). High (10-20 signals) and low (<10 signals) amplification of PPFIA1 was found in 10.9% (5/46) and 21.7% (10/46) tumours, respectively. Fine mapping with overlapping FISH probes showed co-amplification of PPFIA1 and the Cyclin D1 gene which are approximately 600 kb apart from each other, likely in the same amplicon.. PPFIA1 was frequently co-amplified with the Cyclin D1 gene in oral carcinomas and could present a biomarker as well as a novel target for specific gene therapy. Further studies are necessary to investigate the role of PPFIA1 in development and pathogenesis of oral carcinomas.

Topics: Adaptor Proteins, Signal Transducing; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Adhesion Molecules; Chromosomes, Human, Pair 11; Cyclin D1; DNA Probes; DNA, Neoplasm; Female; Gene Amplification; Humans; In Situ Hybridization, Fluorescence; Laryngeal Neoplasms; Male; Mouth Neoplasms; Neoplasm Grading; Neoplasm Staging; Oligonucleotide Array Sequence Analysis; Pharyngeal Neoplasms; Tissue Array Analysis

This study aimed to investigate the expressions and significance of KAI1/CD82 and cyclin D1 in laryngeal squamous cell carcinoma (LSCC).. Real-time quantitative PCR (Q-PCR) and Western blot assay were employed to detect the expressions of KAI1/CD82 and cyclin D1 in the laryngeal tissues of 86 LSCC patients, 32 patients with laryngeal polyp and 38 patients with laryngeal leukoplakia, and the influence of both proteins on the clinicopathological features and survival of LSCC patients.. The changes in mRNA and protein expressions of KAI1/CD82 and cyclin D1 were consistent in three groups, and the expressions of KAI1/CD82 and cyclin D1 were significantly different among three groups (P<0.01 or <0.05). The KAI1/CD82 expression in patients with TNM stage III-IV LSCC, poorly differentiated LSCC, clinical stage III-IV LSCC or lymph node metastasis was markedly lower than that in those with TNM stage I-II LSCC, well differentiated LSCC, clinical stage I-II LSCC or no lymph node metastasis (P<0.01 or <0.05). However, there was no marked difference in KAI1/CD82 expression between males and females and among patients in different age groups (P>0.05). In LSCC patients positive for KAI1/CD82 protein expression, the median survival time was 76 months, which was significantly longer than that in LSCC patients negative for KAI1/CD82 protein expression (48 months; X(2)=16.293, P=0.000). The Cyclin D1 expression in patients with TNM stage III-IV LSCC, poorly differentiated LSCC, or clinical stage III-IV LSCC was dramatically higher than that in patients with TNM stage I-II LSCC, well differentiated LSCC, or clinical stage I-II LSCC (P<0.01 or <0.05). However, no marked difference was noted in cyclin D1 expression between males and females, among patients in different age groups and between patients with and without lymph node metastasis (P>0.05). In LSCC patients positive for cyclin D1 protein expression, the median survival time was 40 months, which was markedly shorter than that in LSCC patients negative for cyclin D1 protein expression (X(2)=9.517, P=0.02). In LSCC patients, there was a negative correlation between KAI1/CD82 expression and cyclin D1 expression (X(2)=7.86, P<0.01).. KAI1/CD82 affects cell cycle. Both KAI1/CD82 and cyclin D1 are involved in the occurrence and development of LSCC, and may provide clinical information for evaluation of invasiveness, metastasis and prognosis of LSCC. Thus, KAI1/CD82 and cyclin D1 may serve as markers for determination of invasiveness, metastasis and prognosis of LSCC.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Chi-Square Distribution; Cyclin D1; Female; Humans; Kangai-1 Protein; Kaplan-Meier Estimate; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Risk Factors; RNA, Messenger; Time Factors

Laryngeal squamous cell carcinoma (LSCC) is a malignant neoplasm exhibiting aggressive phenotype, high recurrence rate, and risk of developing second primary tumors. Current evidence suggests that cells in the invasive front of carcinomas have different molecular profiles compared to those in superficial areas. This study aimed to identify candidate genes in the invasive front and superficial cells from laryngeal carcinomas that would be useful as molecular markers. Invasive front and tumor surface cells of 32 LSCC were evaluated by high-resolution comparative genomic hybridization. Both CCND1 copy number gains and cyclin D1 protein expression were evaluated to confirm gains of 11q13.3. Losses of 3q26.2-q29 and 18q23 were confirmed by loss of heterozygosity analysis. The most frequent chromosomal alterations observed only in invasive front cells involved gains of 1p, 4q, and 9p and losses of 3p, 11p, 12p, 13q, 17q, 18p, 19q, 20q, 21q, and Xp. Gains of 11q13 were detected in both components from glottis and supraglottis but only in invasive front cells from transglottic tumors. Fluorescence in situ hybridization confirmed gains of CCND1/CPE11 in a subset of cases. In supraglottic tumors, cyclin D1 positivity was associated with distant metastasis (P = 0.0018) and with decreased disease-free survival (P = 0.042). Loss of heterozygosity at 3q26.2 and 18q23 were associated with lymph node involvement (P = 0.055) and worsened prognosis, respectively. In conclusion, this study revealed regions that could be targeted in the search for molecular markers in LSCC. Cyclin D1 may be useful as a prognostic marker in supraglottic tumors.

Topics: Carcinoma, Squamous Cell; Chromosome Aberrations; Chromosomes, Human; Comparative Genomic Hybridization; Cyclin D1; Female; Gene Dosage; Humans; Immunohistochemistry; Laryngeal Neoplasms; Laser Capture Microdissection; Loss of Heterozygosity; Lymphatic Metastasis; Male; Microsatellite Instability; Microsatellite Repeats; Middle Aged; Tissue Array Analysis

This study aimed to investigate the expression and significance of ATF-3 in laryngeal squamous cell carcinoma (LSCC).. Expression of ATF-3 was examined using immunohistochemistry methods in samples from 83 cases of LSCC carcinoma. MTT assay was used to detect proliferation of Hep-2 cells after ATF-3 knocked down by siRNA lentivirus. A mouse model was used to investigate the inhibitive role of ATF-3 siRNA in LSCC xenografts. Realtime RCR was used to detect Cyclin D1 expression after ATF-3 downregulation in Hep-2 cells.. The expression of ATF-3 was positively detected in all the 83 cases of LSCC cancer tissues while Only 4 cases of adjacent non-neoplastic tissues were detected with positive ATF-3 expression. The ATF-3 expression was statistically related with T stage, neck nodal metastasis, clinical stage and prognosis of LSCC. Both cell proliferation in vitro and tumor growth in vivo were suppressed after ATF-3 knockdown. Furthermore, the expression of Cyclin D1 was decreased after ATF-3 downregulation in Hep-2 cells.. ATF-3 is involved in the progress of LSCC, and may provide clinical information for evaluation of prognosis of LSCC. The oncologic role of ATF-3 may be correlated with Cyclin D1 regulation.

Topics: Activating Transcription Factor 3; Animals; Carcinoma, Squamous Cell; Cell Line, Tumor; Cyclin D1; Down-Regulation; Female; Gene Expression Regulation, Neoplastic; Humans; Laryngeal Neoplasms; Male; Mice; Prognosis; RNA, Small Interfering; Up-Regulation

Cylin D1(CCDN1) is an important regulator of the cell cycle whose alterations are thought to be involved in cancer development. There have been many studies indicating CCDN1 amplification or over- expression in a variety of cancer types. In addition to gene amplification, the G870A polymorphism may be related with altered CCDN1 activity, and therefore with cancer development. This hypothesis has been tested in different cancer types but results have been contradictory. We therefore aimed to investigate any relationship between CCDN1 A870G genotypes and laryngeal squamous cell cancer development and progression.. A total of 68 Turkish patients with primary laryngeal squamous cell cancer and 133 healthy controls were enrolled. Polymerase chain reaction-restriction fragment length polymorphism analysis was used to determine the CCDN1 genotypes.. No significant association was detected between CCDN1 genotypes and laryngeal squamous cell cancer (LxSCCa) development. Similarly CCDN1 genotypes were not related to clinical parameters of Lx SCCa. However, there was a very significant association between CCDN1 G allele and presence of perineural invasion (p= 0.003; OR: 1.464; CI% 1.073-1.999). CCDN1 G allele frequency was significantly higher in the individuals with perineural invasion (85.7%) when compared to those without (58.5%). The 2 patients who died of disease were both found to possess the GG genotype.. These results pose a controversy in suggesting a protective role of the G allele against LxSCCa development and support the association of CCDN1 gene GG genotype with mortality in patients with LxSCCa.

Topics: Carcinoma, Squamous Cell; Case-Control Studies; Cyclin D1; Female; Follow-Up Studies; Genotype; Humans; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasm Staging; Polymerase Chain Reaction; Polymorphism, Genetic; Prognosis; Survival Rate; Turkey

Various biomarkers might ultimately prove to have prognostic value and could be clinically relevant. It is mandatory confirm the prognostic power of these markers in large, well-designed, and prospective studies.. The aim of this study was to investigate the possible role of specific genes and proteins in laryngeal tumorigenesis.. Genetic analysis by multiple ligation-dependent probe amplification and analysis of protein expression by immunohistochemistry were carried out in a series of 50 tissue samples.. In the smoker normal mucosa group TP53 loss was predominant, whereas in the epithelial precursor lesions (EPLs) CDKN2A loss and BCL2L1 gain were most frequent. EPL with progression presented CTNNB1 loss. Positivity at cytoplasm for β-catenin, cyclin D1 and p53 was detected in all EPL cases with progression to invasive carcinoma. Multivariate analysis showed that expression of β-catenin and loss of CTTNB1 were associated with laryngeal cancer risk.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; bcl-X Protein; beta Catenin; Biomarkers, Tumor; Carcinoma in Situ; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Child; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Disease Progression; Female; Gene Expression Regulation, Neoplastic; Humans; Laryngeal Mucosa; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplastic Stem Cells; Prognosis; Smoking; Tumor Suppressor Protein p53; Young Adult

EGFR (epidermal growth factor receptor), cyclin D1 and Akt/mTOR pathways are active in head and neck cancer. The aim of this study was to explore biomarker expression, their correlations with clinicopathological parameters and their prognostic utility in a cohort of patients with localized squamous laryngeal carcinoma.. We assessed relative messenger RNA expression of EGFR, Akt1, 2, and 3, mTOR and CCND1, copy number variants of the EGFR and CCND1 genes and immunohistochemical protein expression of EGFR, p-Akt308, p-Akt473, pmTOR, PTEN, p53 and cyclin D1 in paraffin-embedded tissue samples of localized laryngeal carcinomas.. In 289 patients with T3-4 (77.8%), node-negative (84.1%) tumors of the larynx, high EGFR and CCND1 mRNA correlated with no or ex-smoking, (p = 0.003 and p = 0.029, respectively), while low Akt3 mRNA correlated with alcohol abuse, N0 stage, total laryngectomy, and absence of neck dissection. At a median follow-up of 74.5 months, high mTOR mRNA expression was marginally associated with shorter disease-free survival (hazard ratio [HR] = 1.54; p = 0.093) and high Akt3 mRNA with shorter overall survival (HR = 1.49; p = 0.0786), in univariate analysis. In multivariate analysis, node-positive status, subglottic-transglottic location, surgery other than total laryngectomy and mTOR/CCND1 mRNA interaction with a hazard ratio of 2.16 (p value for interaction:  0.0010) were independent predictors of relapse, while node-positive status and subglottic-transglottic location were associated with higher risk for death.. In localized laryngeal cancer, clinicopathological parameters and an interaction of high mTOR and CCND1 mRNA expression were found to be associated with poor patient outcome.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Combined Modality Therapy; Cyclin D1; ErbB Receptors; Female; Humans; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Prognosis; Retrospective Studies; RNA, Messenger; TOR Serine-Threonine Kinases

Second primary tumors and recurrences are an important problem in patients with head and neck squamous cell carcinoma. The purpose of this study was to determine the genetic changes in tumor samples to improve knowledge of tumor progression.. Copy number changes of 37 genes were analyzed by multiplex ligation-dependent probe amplification (MLPA) in 36 primary tumors and their corresponding 21 second primary tumors and 15 recurrences.. CCND1 and EMS1 amplifications and gain of BCL2L1 were the most common genetic alterations in the primary tumor, second primary tumor, and recurrence samples. Gains of ERBB2 and PTPN1 were associated with recurrences.. Specific genetic profiles for each group have been found. Similarities between primary tumor and second primary tumor and dissimilarity between primary tumor and recurrence suggest that clinicopathological criteria do not always accurately differentiate these entities. Genetic profiling may aid in the diagnosis and prognosis of these difficult cases.

Topics: Adult; Aged; bcl-X Protein; Carcinoma, Squamous Cell; Cathepsin B; Cell Cycle Proteins; Cortactin; Cross-Sectional Studies; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA-Binding Proteins; Female; Gene Expression Profiling; Humans; Interleukin-18; Lamin Type A; Laryngeal Neoplasms; Male; Membrane Proteins; Middle Aged; Neoplasm Recurrence, Local; Neoplasms, Second Primary; Nucleic Acid Amplification Techniques; Oncogene Proteins; Pharyngeal Neoplasms; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Receptor, ErbB-2; Retrospective Studies; RNA-Binding Proteins; Transcription Factors; Tumor Suppressor Protein p53; Tumor Suppressor Proteins

We recently reported on the identification of the Fas-associated death domain (FADD) as a possible driver of the chromosome 11q13 amplicon and the association between increased FADD expression and disease-specific survival in advanced-stage laryngeal carcinoma. The aim of this study was to examine whether expression of FADD and its Ser194-phosphorylated isoform (pFADD) predicts local control in patients with early-stage glottic carcinoma primarily treated with radiotherapy only.. Immunohistochemical staining for FADD and pFADD was performed on pretreatment biopsy specimens of 92 patients with T1-T2 glottic squamous cell carcinoma primarily treated with radiotherapy between 1996 and 2005. Cox regression analysis was used to correlate expression levels with local control.. High levels of pFADD were associated with significantly better local control (hazard ratio, 2.40; 95% confidence interval, 1.04-5.55; p = 0.040). FADD overexpression showed a trend toward better local control (hazard ratio, 3.656; 95% confidence interval, 0.853-15.663; p = 0.081). Multivariate Cox regression analysis showed that high pFADD expression was the best predictor of local control after radiotherapy.. This study showed that expression of phosphorylated FADD is a new prognostic biomarker for better local control after radiotherapy in patients with early-stage glottic carcinomas.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cortactin; Cyclin D1; Fas-Associated Death Domain Protein; Female; Glottis; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Prognosis; Protein Isoforms

To investigate the effect of Notch1 on cell cycle, apoptosis and migration of laryngeal squamous cell carcinoma cell line Hep-2 and explore its possible molecular mechanism.. Hep-2 cells were divided into three groups: untreated group, empty vector group and Notch1 group. The effects of upregulation of Notch1 expression on cell proliferation, cell cycle and apoptosis were assessed by CCK-8 staining and flow cytometry, respectively, and effect of upregulation of Notch1 expression on cell migration of Hep-2 cells was studied using Boyden chamber assay, and further expression changes of genes related to cell proliferation, cell cycle, apoptosis and migration were detected by semi-quantitative RT-PCR and Western blotting.. Compared with the untreated group and empty vector group, cell proliferation of Hep-2 in the Notch1 group was significantly inhibited (P < 0.05). The results of flow cytometry revealed that the percentage of cells at G0/G1 phase in the Notch1 group was (70.43 +/- 1.36)%, significantly higher than the (46.39 +/- 1.19)% in the untreated group or (48.41 +/- 1.18)% in the empty vector group, and there was a significant difference among the three groups (P = 0.000). In addition, the percentage of apoptotic cells in the Notch1 group was (22.46 +/- 0.90)%, significantly higher than that in the untreated group [(5.77 +/- 0.37)%] or empty vector group [(6.09 +/- 0.20)%], with a significant difference among the three groups (P = 0.000). The results of Boyden chamber assay demonstrated that the number of cells migrated through membrane in the Notch1 group was evidently lower than that in the untreated group and empty vector group. Moreover, the results of semi-quantitative RT-PCR and Western blotting showed that cell proliferation inhibition and cell cycle arrest were closely associated with downregulation of cyclin D1, cyclin E and CDK2 expressions and upregulation of p53 expression. In addition, upregulation of Notch1 expression mediated cell apoptosis was tightly related to upregulation of caspase 3/9 expressions and downregulation of Bcl-2, while the decrease of Hep-2 cell migration ability was obviously correlated with downregulation of MMP-2/-9 expressions.. Notch1 signaling pathway may play a pivotal role in the occurrence and development of laryngeal squamous cell carcinoma. Further study may elucidate that Notchl signaling pathway may become a molecular target for therapy of laryngeal squamous cell carcinoma.

Topics: Apoptosis; Carcinoma, Squamous Cell; Caspase 3; Caspase 9; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase 2; Humans; Laryngeal Neoplasms; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Receptor, Notch1; Signal Transduction; Tumor Suppressor Protein p53

To investigate the prognostic role of antigen KI-67 (Ki-67) and G1/S-specific cyclin-D1 (cyclin-D1) in patients with laryngeal squamous cell carcinoma (LSCC).. Immunohistochemical staining (IHS) was used to determine the protein expression of Ki-67 and cyclin-D1 in LSCC tissues. Kaplan-Meier survival curves was calculated with reference to Ki-67 and cyclin-D1 levels.. Cyclin-D1 and Ki67 were expressed in the nuclei of cancer cells. Among the total of 92 cancer tissues examined by immunohistochemistry, 60 (65.22%) had cyclin-D1 overexpression and 56 (60.87%) had Ki67 overexpression. Cyclin-D1 overexpression is associated with the advanced stage of the cancer (P=0.029), but not with gender, age, stage of cancer, histological differentiation, anatomical site, smoking history and alcohol consumption history. Ki67 overexpression is not associated with the advanced stage, gender, age, histological differentiation, anatomical site, smoking history and alcohol consumption history. A statistically significant correlation was found between lymph node status and the expression of Ki67 (p=0.025). Overexpression of cyclin-D1 was correlated to shorter relapse-free survival period (P<0.001).. Overexpression of cyclin-D1 can be used as a marker to predict relapse in patients with LSCC after primary curative resection.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Female; Follow-Up Studies; G1 Phase; Humans; Immunoenzyme Techniques; Ki-67 Antigen; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; S Phase; Survival Rate

To detect the expression of histone deacetylase 6 (HDAC6) in laryngeal squamous cell carcinoma, and to analyze the effects of downregulation of HDAC6 expression on cell cycle, proliferation and migration of laryngeal squamous cell carcinoma cell line Hep-2 cells, and to explore their possible molecular mechanisms.. Immunohistochemistry was used to detect the expression of HDAC6 protein in 55 cases of laryngeal squamous cell carcinoma and 20 cases of normal laryngeal mucosa. HDAC6 siRNA and control siRNA were transfected into Hep-2 cells via lipofectamine 2000, and the interfering effect was analyzed using Western blotting. The effects of downregulation of HDAC6 expression on cell cycle, proliferation and migration were determined by cell counting kit-8 (CCK-8), flow cytometry and Boyden chamber, respectively. Finally, Western blotting was used to detect the expressions of cell cycle, proliferation and migration related proteins.. There was a high level expression of HDAC6 protein in laryngeal squamous cell carcinoma, and its expression was not related to age and sex of the patients (P > 0.05), but closely associated with the degree of histological differentiation, TNM staging and lymph node metastasis (P < 0.05). HDAC6 siRNA effectively down-regulated the expression of HDAC6 protein in laryngeal squamous cell carcinoma cell line Hep-2 cells, and downregulation of its expression obviously inhibited cell proliferation, arrested cell cycle at G(0)/G(1) phase and decreased cell migration ability in Hep-2 cells. Additionally, the downregulation of HDAC6 protein expression markedly decreased the expressions of cyclin D1, cyclin E, cdk2 and MMP-9 proteins, but increased the expressions of p21 and E-cadherin proteins.. HDAC6 may play a pivotal role in the carcinogenesis and development of laryngeal squamous cell carcinoma. The downregulation of HDAC6 expression-mediated cell proliferation inhibition, cell cycle arrest and decreased cell migration ability may be closely associated with the decrease of cyclin D1, cyclin E, cdk2 and MMP-9 proteins and increase of p21 and E-cadherin proteins.

Topics: Adult; Aged; Antigens, CD; Cadherins; Carcinoma, Squamous Cell; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase 2; Down-Regulation; Female; Histone Deacetylase 6; Histone Deacetylases; Humans; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Matrix Metalloproteinase 9; Middle Aged; Neoplasm Staging; Oncogene Proteins; Proto-Oncogene Proteins p21(ras); RNA, Small Interfering; Transfection

Overexpression of Epidermal Growth Factor Receptor (EGFR) and also of cell cycle control proteins, such as cyclin D1 is a frequent event in squamous cell carcinoma of the larynx (LSSC). Our aim was to correlate their protein levels with telomerase catalytic subunit (h-TERT) expression. Using tissue microarray technology, fifty-five paraffin embedded histologically confirmed primary LSSCs and also ten dysplastic lesions were cored at a diameter of 1.5 mm. Immunohistochemistry (IHC) was performed by the use of anti-EGFR, anti-cyclin D1, and anti-h TERT monoclonal antibodies. Chromogenic in situ hybridization (CISH) analysis was also applied using EGFR gene and chromosome 7 probes, respectively. EGFR, cyclin D1 and h-TERT protein overexpression was observed in 48/55 (87.2%), 19/55 (34.5%) and 21/55 (38.1%) carcinoma cases, respectively. EGFR protein expression was statistically associated with grade (P=0.01), and also with stage (P=0.001) of the examined tumors. Borderline statistical significance was assessed correlating overall cyclin D1 expression to h TERT expression (P=0.06). Simultaneous up regulation of the three proteins was established in 7/55 (12.7%) cases, correlated to the stage of the tumors (P=0.05). EGFR gene amplification was observed in 7/65 (10.7%) carcinomas and dysplasias, whereas chromosome 7 aneuploidy was detected in 4/65 (6.1%) of those cases.Simultaneous up regulation of EGFR, cyclin D1 and h TERT proteins correlates with advanced stage in LSCC. EGFR gene amplification and not only protein over expression maybe is the eligible criterion for targeted therapeutic strategies in those patients.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Chromosomes, Human, Pair 7; Cyclin D1; Enzyme Activation; ErbB Receptors; Female; Humans; Immunohistochemistry; In Situ Hybridization; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Staging; Telomerase; Tissue Array Analysis

Uncontrolled proliferation and a decrease in cell-cell adhesion are one of the most important characteristics of malignancy. Determination of replication-dependent histone H3 can be applied as a proliferative marker. Cyclin D1 (CCND1) regulates the cell cycle by participating in the control of the G1/S phase transition. Claudins (CLDN) are components of tight junctions and may play an essential role in the loss of tissue cohesion. The aim of the study was to assess the mRNA expression of histone H3, cyclin D1, and claudin 7 genes in laryngeal squamous cell carcinoma (LSCC) and adjacent nonneoplastic tissues. The study group consisted of 32 patients with LSCC. Adjacent nonneoplastic tissues of incision lines were used as controls. Quantification of H3, CCND1 and CLDN7 mRNAs was performed by the use of real-time QRT-PCR assay. Molecular analysis showed a significantly higher expression of CCND1 (P = 0.0001; Wilcoxon test) and H3 (P = 0.0141) genes in tumor tissues than in surrounding nonneoplastic tissues. On the contrary, transcriptional activity of claudin 7 gene was higher in histologically normal tissues; however, this difference was not statistically significant (P = 0.1499). The data obtained indicate that laryngeal cancer is characterized by high proliferative potential mediated by increase in cyclin D1 and H3 mRNAs expression.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cell Cycle; Cell Proliferation; Claudins; Cyclin D1; Female; Histones; Humans; Laryngeal Neoplasms; Male; Membrane Proteins; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; Tight Junctions; Transcriptional Activation

Down-regulating human telomerase reverse transcriptase (hTERT) expression will significantly suppress the cell viability of laryngeal squamous cell carcinoma Hep-2, which was mainly due to the inhibition of cyclin D1 and thus G1/S phase transition.. Small-interfering RNA (siRNA) targeting hTERT can arrest the cell cycle of cancer cells, as well as inhibit telomerase activity and cell viability. However, the precise mechanisms still remain unclear. Here, we investigate the regulatory role of hTERT in cyclin D1 in laryngeal squamous carcinoma.. Short hairpin RNAs (shRNAs) specifically targeting hTERT were constructed and expressed in Hep-2 cells. Cell proliferation was measured by CCK-8 assay. Expression of hTERT, cyclin D1, cyclin E, c-myc, and GAPDH was detected by RT-PCR and Western blot; cyclin D1 and hTERT proteins in laryngeal squamous carcinoma tissue microarray were analyzed by quantum dots immunofluorescence.. hTERT silence by shRNAs decreased the proliferation of Hep-2 cells by 76.8% at day 4 (96 h). Furthermore, transfection with hTERT shRNA for 48 h also significantly reduced expression of hTERT, cyclin D1, and c-Myc, but not cyclin E. Quantum dots immunofluorescence analysis of 36 laryngeal squamous carcinoma tissue samples found that hTERT expression was highly correlated with cyclin D1 expression.

Topics: Carcinoma, Squamous Cell; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Cyclin E; Down-Regulation; Gene Expression Regulation, Neoplastic; Humans; Laryngeal Neoplasms; Oncogene Proteins; Proto-Oncogene Proteins c-myc; RNA, Small Interfering; Telomerase

Human papillomavirus (HPV) infection is implicated as an important risk factor in the development of head and neck cancers. Many studies focusing on the relationships between HPV infection and cell cycle proteins immunoexpression in laryngeal lesions have provided contradictory results. The aim of this study was to evaluate the relationships between HPV DNA presence and p16INK4a, p21waf1/cip1, p53 and cyclin D1 immunoexpression in heterogenous HPV-positive and HPV-negative groups of laryngeal cancers and inverted papillomas. The HPV DNA expression was detected using an in situ hybridization method and immunoexpression of p16INK4a, p21waf1/cip1, p53 and cyclin D1 using immunohistochemistry. The immunoexpression of p21waf1/ /cip1 and p53 proteins was lower in the HPV-positive group compared to the HPV-negative group, although only the difference of p53 staining was statistically significant. The immunoexpression of p16INK4a and cyclin D1 was significantly increased in the HPV-positive group compared to the HPV-negative group. The increased immunoexpression of p16INK4a and cyclin D1, and the lower immunoexpression of p21waf1/cip1 and p53 in the HPV-positive group compared to the HPV-negative group, supports the hypothesis that HPV may play an important role in cell cycle dysregulation.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Laryngeal Neoplasms; Middle Aged; Papillomaviridae; Paranasal Sinus Neoplasms; Tumor Suppressor Protein p53

To examine the expression and clinical significance of the Stat3 and Cyclin D1 in laryngeal squamous cell carcinoma (LSCC).. The expression of Stat3 and Cyclin D1 in LSCC tissue were detected by the RT-PCR and Western blot method respectively, and then the relationship between Stat3 and Cyclin D1 protein expression level and relevant clinical factors of LSCC was explored.. Compared to the normal larynx mucosa group, the expression level of Stat3 and Cyclin D1 was significantly higher in LSCC (P < 0.01). The high expression level of Stat3 and Cyclin D1 protein in LSCC was correlated with the clinical stage, tumor differentiation and lymph node metastases (P < 0.01). The expression level of Stat3 was positively correlated with that of Cyclin D1, The correlation coefficient(r) was 0.564 (P < 0.01) for the protein expression and 0.552 (P < 0.01) for the mRNA expression.. Stat3 signaling pathway may play an important role in the tumorigenesis and progression of LSCC. The expression level of Stat3 and its target gene Cyclin D1 can reflect the malignancy degree of LSCC.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Staging; STAT3 Transcription Factor

Ent-11-hydroxy-15-oxo-kaur-16-en-19-oic acid (5F) is known to exhibit antitumor activity, but its mechanism is not completely understood. 5F has not been tested in laryngeal cancer.. Two laryngeal cancer cell lines were treated with 5F. Cell death was analyzed by MTT [3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide] and Annexin V assay. Nuclear factor kappa beta (NF-κB)- and apoptosis-related molecules were examined.. 5F induced laryngeal cancer cell death in a dose-dependent manner. The Annexin V assay and the measurement of cleavage of procaspase-3 and poly(ADP-ribose) polymerase demonstrated that the 5F-induced cell death was mainly apoptotic. 5F slightly reduced the basal level of NF-κB, but significantly suppressed the inducible NF-κB by reducing its transcriptional activity, protecting its inhibitory subunit IκBα from degradation, and suppressing its level in the nucleus. 5F also inhibited pro-proliferative and anti-apoptotic molecules but promoted pro-apoptotic Bax.. 5F induces apoptosis of laryngeal cancer cells by inhibiting NF-κB activation/induction, suppressing pro-proliferative and anti-apoptotic molecules, and promoting pro-apoptotic Bax.

Topics: Apoptosis; bcl-2-Associated X Protein; Blotting, Western; Caspase 3; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Cyclooxygenase 2; Diterpenes; Dose-Response Relationship, Drug; Down-Regulation; Drugs, Chinese Herbal; ErbB Receptors; Humans; I-kappa B Kinase; Inhibitor of Apoptosis Proteins; Laryngeal Neoplasms; Microtubule-Associated Proteins; NF-kappa B; Poly(ADP-ribose) Polymerases; Survivin; Transcription, Genetic; Tumor Necrosis Factor-alpha; Up-Regulation

Head and neck cancer is the sixth most common cancer worldwide and laryngeal cancer represents the largest subgroup. However, the molecular mechanism underlying its malignant behavior and progression is not clarified. Accumulating evidence has shown that Notch1 signaling pathway plays a central role in carcinogenesis, but its potential role in regulating the development of laryngeal carcinoma, has not been characterized. Here, we identified that Notch1 signaling pathway was activated in laryngeal carcinoma accompanied with up-regulation of Notch1 and Hes1 expression. Overexpression of Notch1 in laryngeal carcinoma cell line Hep-2 led to suppression of tumor cellular proliferation and arrested cell cycle in the G0/G1 phase and induced cell apoptosis, which were coupled with the down-regulation of cyclin D1, cyclin E, cdk2 and bcl-2 and up-regulation of caspase-3, caspase-9 and p53. Most importantly, up-regulation of Notch1 expression also reduced the migration of Hep-2 cells, which was closely associated with down-regulation of MMP-2 and MMP-9. The finding may lay a foundation for further investigations into the Notch1 signaling pathway as a potential target for laryngeal carcinoma.

Topics: Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Blotting, Western; Caspase 3; Caspase 9; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase 2; Disease Progression; Flow Cytometry; Gene Expression Regulation, Neoplastic; Homeodomain Proteins; Humans; Laryngeal Neoplasms; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Neoplasms, Squamous Cell; Proto-Oncogene Proteins c-bcl-2; Receptor, Notch1; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Transcription Factor HES-1; Transfection; Tumor Suppressor Protein p53

To study the expression and relationship of Pin1 and CyclinD1 in adult papilloma of larynx, and the effect of both in laryngeal papilloma's canceration.. Ninety-two cases of paraffin section with immunoperoxidase (SP) staining method was used to detect the distribution of Pin1 and CyclinD1 in 10 cases of laryngeal normal epithelial tissue, 39 cases of laryngeal papilloma, 27 cases of laryngeal papilloma with middle, severe atypical hyperplasia and 16 cases of laryngeal carcinoma.. The distribution of Pin1 and CyclinD1 increased gradually from laryngeal normal epithelial tissue to laryngeal carcinoma (P<0.05); No difference of the expression of CyclinD1 (not including Pin1, for Pin1, P=0.009) was found between laryngeal papilloma and laryngeal papilloma with middle, severe atypical hyperplasia (P>0.0125), but there had significant difference of the expression of Pin1 and CyclinD1 among the rest groups; There was significantly direct correlation between the expression of Pin1 and CyclinD1 (P<0.05).. The hyper-expressions of Pin1 and CyclinD1 may play a key role in laryngeal papilloma's malignant change. Pin1 up-regulating the expressions of cyclinD1 possibly participate in its malignant change.

Topics: Adult; Aged; Cyclin D1; Female; Humans; Laryngeal Neoplasms; Male; Middle Aged; NIMA-Interacting Peptidylprolyl Isomerase; Papilloma; Peptidylprolyl Isomerase; Young Adult

To investigate the overexpression of Stat3 and Cyclin D1 in laryngeal neoplasm with immunohistochemistry method.. With immunohistochemistry method, we investigated the expression of Stat3 and Cyclin D1 in laryngeal neoplasm and analysis the relationship between Stat3 and clinical pathological factor.. Stat3 and Cyclin D1 overexpressed in laryngeal neoplasm tissue and they have positive relationship.. Active Stat3 may promote the transcription of target gene Cyclin D1, which could accelerate carcinomatous change.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Staging; STAT3 Transcription Factor

The commercially available analogue of indirubin, indirubin-3-oxime, is known for its antitumor activities. To further establish its role in anticancer activity, we tested its potential against human laryngeal carcinoma cell line (Hep-2). We investigated the molecular mechanisms of indirubin-induced apoptosis and growth arrest in human laryngeal carcinoma cells. Upon treatment with indirubin-3-monooxime, a time dependent inhibition of cell growth was observed and cells developed many hallmark features of apoptosis. The increase of chromatin condensation after treatment with indirubin-3-oxime identified by staining with chromatin stain Hoechst 333258 indicates apoptosis. The observed increase in DNA fragmentation after indirubin-3-oxime in DNA gel electrophoresis indicates the apoptotic feature exhibited by the drug. Flow cytometric analysis confirmed that indirubin increased populations of apoptotic phase G1. Indirubin-3-oxime-induced growth inhibition was associated with induction of Cdk inhibitor p21, inhibition of cyclin D1 and activation of caspase-3. We conclude that indirubin-3-oxime induces cell death and apoptosis in human laryngeal carcinoma cells.

Topics: Antineoplastic Agents; Apoptosis; Caspase 3; Cell Cycle; Cell Line, Tumor; Chromatin; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; DNA Fragmentation; Drug Screening Assays, Antitumor; Electrophoresis, Agar Gel; Flow Cytometry; Humans; Indoles; Laryngeal Neoplasms; Oximes; Time Factors

Constitutive activation of signal transducers and activators of transcription (STAT) 3 has been observed in many solid tumors including head and neck squamous cell carcinoma. Expression and activation of STAT3 in laryngeal carcinoma have not been fully understood. The study aims to investigate the expression and activation of STAT3 in laryngeal carcinoma, the relationship between activated STAT3 and its downstream target gene CyclinD1 and the related clinicopathological factors of activated STAT3.. Prospective.. Sixty-four samples of laryngeal squamous cell carcinoma and 12 samples of control mucosa obtained from total laryngectomy cases were analyzed using Western blot analysis and reverse transcriptase-polymerase chain reaction. Statistical analysis was performed using SPSS.. The overexpression of both STAT3 and CyclinD1 mRNA was observed in all samples of laryngeal squamous cell carcinoma. The mRNA levels of STAT3 and CyclinD1 in carcinoma tissue were 2.1- and 2.3-fold higher than those in control mucosa, respectively; the differences were statistically significant (P < .01). The overexpression of STAT3, p-STAT3, and CyclinD1 protein was also observed in all tumor samples. The protein levels of STAT3, p-STAT3, and CyclinD1 in carcinoma tissue were 1.6-, 4.5-, and 2.0-fold higher than those in control mucosa respectively; the differences were statistically significant (P < .01). There was a positive correlation between p-STAT3 protein and CyclinD1 mRNA (Pearson correlation coefficient = 0.827, P < .01). There were significant correlations between the overexpression of p-STAT3 protein and clinical T stage (P < .01), and tumor size (P < .05). The p-STAT3 protein level of patients in T1, T2 was higher than that of patients in T3, T4. The p-STAT3 protein level of patients with tumor size within 20 mm was higher than that of patients with tumor size more than 20 mm.. High expression and activation of STAT3 exist in laryngeal carcinomas. Activated STAT3 may take effect on promoting transcription of its downstream target gene CyclinD1. The role of activation of STAT3 in laryngeal carcinogenesis needs further research.

Topics: Blotting, Western; Carcinoma, Squamous Cell; Cyclin D1; Disease Progression; Female; Gene Expression Regulation, Neoplastic; Gene Targeting; Humans; Laryngeal Neoplasms; Male; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm; STAT3 Transcription Factor; Transcriptional Activation

CyclinD1 and p16 are involved in the regulation of G1 checkpoint and may play an important role in the tumorgenesis of laryngeal squamous cell carcinoma (LSCC). Previous studies have examined the level of expression of cyclinD1 or p16 in LSCC but no such information is available for their relation and their correlation with lymph node metastasis in Chinese patients.. A total of 58 patients underwent surgical resection of laryngeal tumours in the Department of Otolaryngology, Qilu Hospital Shandong University between January 2001 and December 2002. All pathologic specimens were available for immunohistochemical study using antibodies against cyclinD1 and p16.. Compared with normal epithelium, expression of CyclinD1 in the LSCC was significantly higher (62.1% vs. 10.0%, P < 0.05), expression of p16 was significantly lower (48.3% vs. 90.0%, P < 0.05); CyclinD1 expression in LSCC was up-regulated in TNM classification (r (s) = 0.409, P < 0.05) as well as with cervical lymph node metastases (r (s) = 0.294, P < 0.05); p16 expression in LSCC was down-regulated with cervical lymph node metastases (r (s) = -0.275, P < 0.05); negative significant correlation between p16 immunostaining and CyclinD1 was observed in LSCC (r (s) = -0.331, P < 0.05).. There was a negative correlation between CyclinD1 expression and p16 expression in LSCC. The over-expression of CyclinD1 and the under-expression of p16 may play a significant role in the incidence and development of LSCC and may be important indicators for cervical lymph node metastases in Chinese patients of LSCC.

Topics: Adult; Aged; Carcinoma, Squamous Cell; China; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Humans; Immunoenzyme Techniques; Laryngeal Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Prognosis

To investigate the factors of laryngeal carcinoma recurrence, 103 patients of laryngeal carcinoma were analyzed retrospectively on carcinoma marker, molecular margin and clinical factors with univariate analysis and multivariate analysis.. CyclinD1, p27, p53 and eIF4E in primary site and surgery margins were detected in laryngeal carcinoma recurrence group and unrecurrence group with immunohistochemical staining to explore the significance of CyclinD1, p27, p53 and eIF4E on laryngeal carcinoma recurrence; The clinical data of 103 patients of laryngeal carcinoma were analyzed retrospectively to investigate the clinical factors of laryngeal carcinoma recurrence; At last above three factors were analyzed with multivariate analysis.. There was significant difference between laryngeal carcinoma recurrence group and unrecurrence group about CyclinD1, p27 and p53 in laryngeal primary site; There was no significant difference between laryngeal carcinoma recurrence group and unrecurrence group about eIF4E. There was significant difference between laryngeal carcinoma recurrence group and unrecurrence group about CyclinD1, p27, p53 and eIF4E in surgery margins. Laryngeal carcinoma recurrence after surgery was related with carcinoma site, T stage, node metastasis, laryngeal carcinoma pathology and operative method; However, it was not related with age, sex and postoperative irradiation therapy with univariate analysis. Laryngeal carcinoma recurrence after surgery was related with T stage, node metastasis, laryngeal carcinoma pathology and operative method with logistic multivariate analysis. At last, laryngeal carcinoma recurrence after surgery was related with T stage, node metastasis, laryngeal carcinoma pathology and positive molecular margins with logistic multivariate analysis.. The factors of laryngeal carcinoma recurrence is comprehensive. T stage, node metastasis,laryngeal carcinoma pathology and laryngeal carcinoma positive molecular margins were related with laryngeal carcinoma recurrence. Positive molecular margins were more reliable.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p27; Eukaryotic Initiation Factor-4E; Female; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; Tumor Suppressor Protein p53

Amplification of the 11q13 region is one of the most frequent aberrations in squamous cell carcinomas of the head and neck region (HNSCC). Amplification of 11q13 has been shown to correlate with the presence of lymph node metastases and decreased survival. The 11q13.3 amplicon carries numerous genes including cyclin D1 and cortactin. Recently, we reported that FADD becomes overexpressed upon amplification and that FADD protein expression predicts for lymph node positivity and disease-specific mortality. However, the gene within the 11q13.3 amplicon responsible for this correlation is yet to be identified. In this paper, we compared, using immunohistochemical analysis for cyclin D1, FADD and cortactin in a series of 106 laryngeal carcinomas which gene correlates best with lymph node metastases and increased disease-specific mortality. Univariate Cox regression analysis revealed that high expression of cyclin D1 (P=0.016), FADD (P=0.003) and cortactin (P=0.0006) predict for increased risk to disease-specific mortality. Multivariate Cox analysis revealed that only high cortactin expression correlates with disease-specific mortality independent of cyclin D1 and/or FADD. Of genes located in the 11q13 amplicon, cortactin expression is the best predictor for shorter disease-specific survival in late stage laryngeal carcinomas.

Topics: Adult; Aged; Aged, 80 and over; Chromosomes, Human, Pair 11; Cortactin; Cyclin D1; Fas-Associated Death Domain Protein; Female; Gene Amplification; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Staging; Regression Analysis

To evaluate the prognostic value of the expressions of epidermal growth factor receptor (EGFR) and cyclin D1 in early glottic cancer treated with radiotherapy only.. One hundred fifty-one patients with T1-2, N0 glottic cancer who had been treated with radiotherapy at Seoul National University Hospital since 1992 through 2004. Immunohistochemical staining for EGFR and cyclin D1 were performed on the formalin-fixed paraffin-embedded tissues of 25 patients who developed local recurrence and on the tissues of 25 matched patients free from disease. Patterns and degrees of expression were compared between these 2 groups.. High EGFR (p = .047) and high cyclin D1 (p = .040) expressions were both found to be significantly associated with a poor prognosis. No association was found between EGFR and cyclin D1 status (p = .158), but EGFR and cyclin D1 status in combination were found to be significantly associated with local control. The patients with both high EGFR and high cyclin D1 expression had the poorest outcome compared with the others (14 months vs 29 months of median time to progression). Patterns of EGFR and cyclin D1 expression changed after recurrence, but these changes were not found to alter the ultimate prognosis.. The molecular biomarkers, EGFR and cyclin D1 have a prognostic significance in early glottic cancer. These markers in combination seem to play an important role in tumor relapse and may be useful for selecting patients with a poor outcome after radiotherapy.

Topics: Biomarkers, Tumor; Biopsy, Needle; Carcinoma, Squamous Cell; Case-Control Studies; Cyclin D1; Early Diagnosis; Epidermal Growth Factor; Female; Follow-Up Studies; Glottis; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Neoadjuvant Therapy; Neoplasm Recurrence, Local; Neoplasm Staging; Probability; Reference Values; Retrospective Studies; Risk Assessment; Survival Analysis; Treatment Outcome

Cell cycle modulators are important in carcinogenesis and may be of prognostic and therapeutic relevance. This study has examined the influence of the proliferation index (Ki-67) and immunocytochemical expression of epidermal growth factor receptor (EGFR), cyclin D1, and retinoblastoma protein on recurrence rates at the primary site in 50 patients with T2N0 laryngeal carcinomas treated with radical irradiation. Pre-treatment biopsies were retrieved and sections scored for the four immunocytochemical markers. Statistical analysis for association, interaction and survival was performed. Five cases showed loss of expression of Rb protein. The median Ki-67 index was 50%, the median cyclin D1 index 21% and the median EGFR index 47% of cells. EGFR and cyclin D1 expression were positively correlated and, whilst local recurrence tended to occur with a Ki-67 labelling index of <50%, this was not statistically significant. When interactions were examined using Multiple Logistic Regression it was found that there was a direct relationship between EGFR and cyclin D1 expression. If the EGFR index was >20% and the cyclin D1 index >10%, then the odds ratio in favour of a primary site recurrence was very high (5.32 +/-0.41). This study demonstrates that the relationship between EGFR index and cyclin D1 index has a very strong association with primary site recurrence for T2 N0 laryngeal carcinomas treated by irradiation.

Topics: Aged; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cyclin D1; ErbB Receptors; Female; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Odds Ratio; Retinoblastoma Protein; Survival Analysis

Contrary to most reports, our study shows that the expression of cyclin D1 is not an independent prognostic factor in patients with laryngeal cancer (LC). No correlation between cyclin D1 expression and survival rates in LC was found in a multivariate analysis.. The aim of this study was to determine the possible relevance of the expression of cyclin D1 protein in LC as prognostic criteria and to analyse correlation of the expression with clinicopathological features and survival rates.. Immunohistochemistry staining was used to detect the expression of cyclin D1 in 130 samples of laryngeal cancer and in 22 specimens of laryngeal nodules.. Cyclin D1 expression was found in 52 (40%) LC samples and in 3 (13.6%) samples of laryngeal nodules. There was no significant correlation between cyclin D1 expression and clinicopathological features of LC. A multivariate analysis of survival confirmed that cyclin D1 expression was not an independent prognostic factor in LC.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Cyclin D1; Female; Follow-Up Studies; Humans; Immunohistochemistry; Laryngeal Mucosa; Laryngeal Neoplasms; Male; Middle Aged; Multivariate Analysis; Neoplasm Recurrence, Local; Prognosis; Survival Analysis

Cyclin D1 is one of the key proteins involved in cell cycle control, and it is believed that its overexpression may be connected with tumorigenesis. A reason for cyclin D1 deregulation may be connected to a common G870A polymorphism at codon 242 in exon 4 of the CCND1 gene. This single nucleotide substitution, localized in the conserved splice donor site between exon 4 and the intron 4 boundary, might modulate the frequency of alternative splicing. It has been postulated that the A allele results in a higher level of mRNA (transcript b) encoding a protein with an altered C-terminal domain. The influence of CCND1 G-->A polymorphism for the risk of cancer and the prognosis of patients with different types of solid tumors has already been suggested. This study was conducted to investigate the association between the cyclin D1 gene polymorphism and laryngeal cancer risk, as well as the clinical outcome. We also examined the relationship between genotype/allele distributions and the cyclin D1 expression profile. The genotyping study was done using the PCR-RFLP method in 63 patients with larynx cancer and 102 healthy controls. The heterozygotic genotype GA as well as a combination of GA and AA genotypes were associated with an increased risk of larynx cancer compared to the GG genotype (OR =3.02; P =0.004 and OR =2.52; P =0.013, respectively). The A allele frequency was higher in cancer cases (0.484) than in controls (0.416) that were connected with a slightly increased risk of cancer development (OR =1.34); however, the difference was not significant. The AA genotype was associated with an early cancer onset compared to the GG genotype (median age: 51.5 and 63.0 years, respectively). We also demonstrated that the AA genotype was associated with the occurrence of lymph node metastases (OR =3.26) and a higher level of cyclin D1 overexpression. These results suggest that the CCND1 A allele may be a genetic factor that modulates the risk of larynx cancer development, and it may also have an effect on tumor biology and disease prognosis.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cyclin D1; Female; Gene Frequency; Genes, bcl-1; Genetic Predisposition to Disease; Genotype; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide

To study the effect of pSilencer1.0-U6-siRNA-stat3 on the growth of human laryngeal tumors in nude mice.. Hep2 cells were transplanted into nude mice, then at the time of tumor formation, growth rates were observed. After the tumor formed, pSilencer1.0-U6-siRNA-stat3 was injected. Tumor volumes were calculated, and growth curves were plotted. Representative histological sections were taken from mice bearing transplantation tumors in both treated and control groups, and stat3, pTyr-stat3, Bcl-2, cyclin D1, and survivin expression were detected by Western blotting. survivin mRNA levels were detected by Northern blotting, hematoxylin and eosin staining and terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) assay to confirm the apoptosis of tumors.. In nude mice, pSilencer1.0-U6-siRNA-stat3 significantly suppressed the growth of tumors compared with controls (P<0.01). It suppressed stat3 expression, and downregulated BcL2, cyclin D1, and survivin expression within the tumor. This significantly induced apoptosis of the tumors.. pSilencer1.0-U6-siRNA-stat3 was able to inhibit the growth of transplanted human laryngeal tumors in nude mice and induce apoptosis.

Topics: Animals; Apoptosis; Cell Line, Tumor; Cyclin D1; Humans; Inhibitor of Apoptosis Proteins; Laryngeal Neoplasms; Mice; Mice, Nude; Microtubule-Associated Proteins; Neoplasm Proteins; Neoplasm Transplantation; Proto-Oncogene Proteins c-bcl-2; RNA Interference; RNA, Messenger; RNA, Small Interfering; STAT3 Transcription Factor; Survivin

To investigate the response of tumour growth to cisplatin treatment, in relation to p53 mutation and cyclin D1 dysregulation on DNA and protein level, biopsies from seven xenografted human squamous cell carcinomas from the head and neck were analysed with immunohistochemistry for p53 expression and cyclin D1 expression. Polymerase chain reaction-singlestranded conformation polymorphism was used to determine p53 mutations. Fluorescence in situ hybridization was performed to analyse cyclin D1 amplification. The mice were injected i.p. with NaCl (controls) or cisplatin. After injection the tumour volume were measured. The inhibition of tumour growth by cisplatin was defined as the area under the growth curves, and compared with the growth curves of the tumours in the control group. Xenografts with p53 mutation showed significantly higher resistance to cisplatin (p < 0.001) and also tumours with cyclin D1 amplification showed significantly higher resistance (p < 0.001).

Topics: Animals; Carcinoma, Squamous Cell; Cheek; Cisplatin; Cyclin D1; Drug Resistance, Neoplasm; Flow Cytometry; Gene Amplification; Gingival Neoplasms; Head and Neck Neoplasms; Humans; Laryngeal Neoplasms; Mice; Mouth Neoplasms; Mutation; Neoplasms, Unknown Primary; Statistics as Topic; Tumor Suppressor Protein p53; Xenograft Model Antitumor Assays

In this paper we carried out an immunohistochemical study of cyclin D1 (DCS6 ) expression in a series of 195 patients with laryngeal carcinoma that were diagnosticated, treated and followed at the Department of Otolaryngology at "Virgen de la Salud" Hospital (Toledo, Spain) for a time of 5 years. In the cases with lymph node metastasis we also studied cyclin D1 expression at this level. Furthermore we have analysed the value of cyclin D1 expression as a prognostic factor (tumor recurrence, deads due to cancer and survival) and we evaluate the relationship between cyclin D1 expression and other clinic and pathologic parameters.

Topics: Aged; Carcinoma, Squamous Cell; Cyclin D1; Humans; Immunohistochemistry; Laryngeal Neoplasms; Prognosis

Chromosomal aberrations were analyzed in 12 established cell lines derived from laryngeal squamous cell carcinoma. Cytogenetic and fluorescence in situ hybridization studies were used to identify aberrations in the 11q13 region and in some other chromosome regions. Amplification of 11q13 was established only in the cell lines derived from subjects with a survival period of less than 5 years and, together with the 3q gain, were the only chromosomal structural abnormalities connected with short survival. In this group we also found translocations with a breakpoint within 11q13. In three cell lines, 11q13 was observed as a homogenously staining region. The results suggest that amplification of 11q13, as well as re-arrangements potentially involved in up-regulation of the oncogenes mapped in 11q13, should be considered as markers of poor prognosis in laryngeal cancer. A diagnostic significance of 11q13 may be increased by a parallel determination with 3q gain.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Line, Tumor; Chromosome Aberrations; Chromosomes, Human, Pair 11; Cyclin D1; Humans; Karyotyping; Laryngeal Neoplasms; Male; Middle Aged; Prognosis

The prognostic divergence of laryngeal and hypopharyngeal carcinomas is well known. Hypopharyngeal tumors are characterized by frequent metastasis formation and local recurrence, which is the source of the unfavorable prognosis of this subtype. The aim of this study was to define chromosomal alterations associated with the aggressive behavior of hypopharyngeal tumors.. Twenty-nine head and neck squamous cell carcinomas (larynx n = 14 and hypopharynx n = 15) were analyzed by comparative genomic hybridization (CGH). Fluorescence in situ hybridization (FISH) was used to validate the CGH data and to compare the amplification pattern of the most frequently altered gene (cyclin-D1, CCND1) located on 11q13.. The average number of genetic alterations was significantly higher in the hypopharyngeal tumors (P = 0.02). A good correlation of FISH and CGH data were seen. Gains on 11q13 were present in both subtypes, whereas amplification of CCND1 was associated with the aggressive phenotype by FISH. Chromosomal alteration, which was rarely detected in hypopharyngeal tumors but was observed in more than 50% of laryngeal carcinomas, was 8q gain.. Our CGH and FISH data show that head and neck squamous cell carcinomas contain complex cytogenetic alterations and further support the hypothesis that different molecular pathways are responsible for the progression of differently localized tumors of the upper aerodigestive tract.

Topics: Aged; Chromosome Aberrations; Chromosomes, Human, Pair 11; Cyclin D1; Female; Gene Dosage; Humans; Hypopharyngeal Neoplasms; In Situ Hybridization, Fluorescence; Laryngeal Neoplasms; Male; Middle Aged; Nucleic Acid Hybridization

Cyclin D1 (CCND1) is a set of periodic regulatory proteins that is believed to govern cell cycle transit from G1 into S phase. Overexpression of CCND1 leads to abnormal cellular proliferation which underlies processes of tumorigenesis; CCND1 can thus function as a cooperative oncogene in cell transformation. In the present study we investigate the immunohistochemical expression of CCND1 in a well-documented series of 58 laryngeal squamous cell carcinomas (LSCC) and search for statistical associations between CCND1 index and various clinicopathological parameters including several immunomarkers' expression as well as patients' disease-free survival. Tissue sections from archival paraffin blocks were stained using the avidin-biotin-peroxidase complex method; the H-295 rabbit polyclonal antibody was applied at dilution of 1:150. The percentage of CCND1 immunoreactive tumor cells for each tumor was counted by an image analysis system. CCND1 staining was confined to cell nuclei and, in the examined samples, ranged from undetectable (i.e. 0% of tumor cells, n = 6) to the majority of tumor cells (i.e. 89% of tumor cells) with mean value: 15.73%. In tumor adjacent, non invasive lesions, strong CCND1 staining was noticed in areas with cellular atypia. In cases with nodal metastases, no change in CCND1 expression in the nodal metastases compared with the primary tumors was observed. p53 protein accumulation in malignant cells was positively linked with CCND1 index (Mann-Whitney U: 205.5, p = 0.034). CCND1 expression appears to be an early event in processes of tumorigenesis and tumor progression in some LSCC. Apart from p53 protein accumulation, CCND1 immunohistochemical expression does not seem to correlate with nodal metastasis, disease recurrence or any other clinicopathological prognostic indicator.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Chromosomes, Human, Pair 11; Cyclin D1; Female; Follow-Up Studies; Genes, p53; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Staging; Recurrence

The aim of this study was to evaluate correlations between DNA ploidy type, the immunostaining of cyclin D1, bcl-2 and the intensity of lymphocytic infiltration of the tumor microenvironment in relation to the histopathological G differentiation and pTNM classification. Thirty two patients were treated surgically for laryngeal cancer with total or partial laryngectomy in the Department of Otolaryngology Zabrze. The percentage of bcl-2 immunostaining was showed in 53% of the cases and was found to correlate with G differentiation and the patients' age. Cyclin D1 antigen stained positively in 24 cases (75%). Expression of cyclin D1 correlated with cancer stage. Cyclin D1 negative stain was found in T4-stage. DNA ploidy was examined in 19 cases. Aneuploidy was found in 5 cases only, while the rest were diploid. DNA ploidy value correlated with cyclin D1 expression. All paraffin sections were found to contain lymphocytic infiltrations of CD43 and CD45RO phenotype in the tumor front. Some cases showed high intensity of lymphocytic infiltrations of CD45RO phenotype. The intensity of CD43 lymphocytic infiltrations in the tumor front was related to the expression of cyclin D1 and bcl-2.

Topics: Adult; Aged; Aneuploidy; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; DNA, Neoplasm; Female; Humans; Laryngeal Neoplasms; Laryngectomy; Lymphocytes, Tumor-Infiltrating; Male; Middle Aged; Prognosis; Proto-Oncogene Proteins c-bcl-2

Squamous cell carcinoma of the upper aerodigestive tract (UADT) is associated with environmental factors, especially tobacco and alcohol consumption. Genetic factors, including cyclin D1 (CCND1) polymorphism have been suggested to play an important role in tumorigenesis and progression of UADT cancer. To investigate the relationship between CCND1 polymorphism on susceptibility for UADT cancers, 147 cancer and 135 non-cancer subjects were included in this study. CCND1 genotype at codon 242(G870A) in exon 4 was undertaken using denaturing high performance liquid chromatography (DHPLC) and DNA sequencing. Significant odds ratio (OR) of the AA+GA genotypes [OR=7.5 (95% CI: 1.4-39.7)] was observed in non-drinkers but for non-smokers a non-significant [OR=5.4 (95% CI: 0.9-31.4)] was found in the adjusted model. These results suggest that allele A may be a risk factor for UADT cancer, especially in non-alcoholics. However, further epidemiological studies are needed to establish the exact role of CCND1 polymorphism and the development of UADT cancers.

Topics: Adult; Age Factors; Aged; Aged, 80 and over; Alleles; Carcinoma, Squamous Cell; Cyclin D1; Female; Genotype; Head and Neck Neoplasms; Humans; Hypopharyngeal Neoplasms; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Oropharyngeal Neoplasms; Polymorphism, Genetic; Risk Factors

Altered Cyclin D1 activity, due to gene amplification and/or protein overexpression, is related to the development of several human cancers, including head and neck SCC. This study investigated the relationship between CCND1 A870G gene polymorphism and amplification with the development and progression of laryngeal SCC, considering the implications of tumor localization and tobacco exposure. The study population consisted of 66 larynx cancer patients and 110 healthy individuals. CCND1 A/G polymorphism in exon 4 was genotyped by a PCR-RFLP assay. Cyclin D1 gene amplification was evaluated by a Differential-PCR assay and determined by a quantitative densitometric analysis. Our data on gene amplification did not show any correlation with disease stage, histological tumor differentiation, recurrent disease, disease-specific survival or tumor location. However, GG870 genotype was associated with a shorter disease free interval and a reduced overall survival in laryngeal cancer patients. Moreover, this constitutes the first report of a correlation between cyclin D1 A870G polymorphism and increased susceptibility for laryngeal tumor development at the glottic region, which supports the theory of site-specific prevalence of genetic alterations.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Case-Control Studies; Cyclin D1; Disease Progression; Disease-Free Survival; Female; Gene Amplification; Genotype; Humans; Laryngeal Neoplasms; Male; Middle Aged; Polymerase Chain Reaction; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Prognosis; Tobacco Smoke Pollution

The expression of cell-cycle progression molecules cyclin D1 and cyclin E were immunohistochemically examined in a series of 64 squamous cell invasive carcinomas of the larynx, 10 in situ carcinomas, 34 cases of dysplasia, 11 papillomas and 23 cases of keratosis. The results of their expression were compared with two cell-cycle implicated tumor suppressor proteins p53 and pRb as well as with two proliferation associated indices PCNA and Ki-67 in an attempt to elucidate their potential role in the pathogenesis and progression of these lesions. Nuclear staining for cyclin D1 and E (>5% positive cells) was observed in 19% and 39.7% of the laryngeal carcinomas, respectively. Significantly elevated levels of cyclin D1 and E in invasive laryngeal carcinomas compared with in situ carcinomas were revealed (p=0.045 and p=0.0003, respectively). High levels of cyclin D1 and E expression were correlated with increased Ki-67 score (p=0.037 and 0.017 respectively). A significant positive correlation between cyclin D1 and E was also detected in carcinomas (p=0.018). Decreased levels of cyclins D1 and E in the group of in situ carcinomas compared with those of dysplastic cases and papillomas were also observed. In the dysplastic lesions cyclin D1 expression was correlated with pRb expression (p=0.02). In the cases of keratosis cyclins D1 and E expression were correlated with pRb (p=0.002 and p=0.036, respectively), while cyclin D1 was associated with PCNA (p=0.008) and Ki-67 score (p=0.009). The prognostic significance of cyclins D1, E in determining the risk of recurrence and overall survival with both univariate (long-rang test) and multivariate (Cox regression) methods of analysis showed no statistically significant differences. We conclude that the expression of cyclins D1 and E in squamous cell carcinomas of the larynx does not seem to have a prognostic significance. In addition, their expression may be involved in the development of laryngeal lesions, implicated in cell proliferation, with other cell cycle related proteins, probably by different molecular pathways.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma in Situ; Carcinoma, Squamous Cell; Cell Cycle; Cohort Studies; Cyclin D1; Cyclin E; Female; Humans; Immunoenzyme Techniques; Keratosis; Ki-67 Antigen; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Neoplasms, Glandular and Epithelial; Papilloma; Proliferating Cell Nuclear Antigen; Retinoblastoma Protein; Tumor Suppressor Protein p53

We sought to study various parameters in laryngeal squamous cell carcinoma (LSCC) that might predict nodal metastasis.. Sixty-four LSCCs were examined with respect to their histopathology and, using immunohistochemistry, their proliferative capacity (MIB1), p53 and cyclin D1 status, and intratumoral microvessel density. The presence of human papillomavirus was ascertained by the polymerase chain reaction.. Histopathologically, most tumors had an infiltrating/mixed growth pattern and a diminished inflammatory reaction at the growing margin. In addition, 56% of the tumors were positive for MIB1, with 64% showing p53 overexpression; 70% were positive for cyclin D1; and 59% showed increased tumor microvessel density. Of 42 cases analyzed, 9.5% were positive for human papillomavirus 16.. Of the parameters studied, a diminished lymphocytic inflammatory response at the periphery (P < 0.05) and cyclin D1 overexpression (P < 0.001) correlated significantly with cervical nodal metastasis at presentation.. Cyclin D1 overexpression, easily assessed on biopsy samples, may thus help in optimizing therapy at the outset.

Topics: Adolescent; Adult; Carcinoma, Squamous Cell; Cyclin D1; DNA-Binding Proteins; Female; Humans; Immunohistochemistry; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Papillomaviridae; Polymerase Chain Reaction; Transcription Factors; Tumor Suppressor Protein p53

Defects in the system controlling the cell cycle can lead to an increased proliferation of cancer cells. The aim of this study was to analyse the immunohistochemical expression of chosen cell cycle proteins (P16, cyclin D1 and retinoblastoma protein) and their connection with the clinical course of the disease in laryngeal squamous cell cancer (LSCC). Cancer tissue sections obtained from 58 patients after total laryngectomy served to determine the level of the proteins' expression using immunohistochemical staining and commercial antibodies. A decreased level of P16 expression in 47 per cent, of retinoblastoma protein in 12 per cent and strong cyclin D1 expression in 48 per cent of cases was revealed. Our results show significant correlation between decreased P16 expression and increased tumour dedifferentiation. Overexpression of cyclin D1 was statistically more common in locally advanced tumours (T(3) -T(4)). Low expression of retinoblastoma protein was significantly correlated with both positive P16 immunostaining and with strong cyclin D1 expression. Our study confirms that dysfunction of cell cycle regulation is a common event and may play a significant role in the development of squamous cell carcinoma of the larynx.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cell Differentiation; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Proteins; Prognosis; Retinoblastoma Protein

Tumour cells are characterised by uncontrolled growth due to alterations in the genes that play a key role in cell repair systems and apoptosis: pro-mitotic oncogenes such as cyclin D1, and tumour suppressor genes such as p27. Recent studies have demonstrated that these genes are involved in different epithelial neoplasms and that their expression is generally associated with prognosis. The aim of this immunohistochemical study was to analyse the clinical relevance of cyclin D1/p27 co-expression in a homogeneous series of 132 laryngeal squamous cell carcinomas. Multivariate analysis showed that cyclin D1 and p27 were the only statistically significant predictors of disease-free and overall survival. In relation to the simultaneous expression of p27 protein and cyclin D1, the patients with a cyclin D1+/p27-phenotype had the poorest disease-free and overall survival rates. On the basis of these immunohistochemical results, it was possible to select a subgroup of patients with a high risk of recurrence and poor prognosis to undergo more extended surgical treatment and/or combination antitumoral therapeutic procedures.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Cycle; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p27; Female; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Prognosis; Survival Rate; Tumor Suppressor Proteins

Proteins p53 and cyclin D1 play a crucial role in cell cycle control. Protein p53 mutations are one of the most common genetic alterations in human cancer, and cyclin D1 gene amplification has been found to be associated with poor prognosis in different types of tumors. Functional alterations of these proteins may play an important role both in the carcinogenesis of squamous carcinomas of the head and neck and in the clinical evolution of these tumors. The objective of the present study was to evaluate whether the presence of p53 and/or cyclin D1 proteins (detected by immunohistochemical analysis) could serve as relevant variables for the assessment of the prognosis of laryngeal squamous cell carcinoma.. Retrospective multicentric study.. Paraffin-embedded biopsy specimens from 62 human epidermoid laryngeal carcinomas were randomly selected. The expression of p53 and cyclin D1 was examined by means of immunohistochemical analysis with a view to evaluating whether there is a correlation between the aberrant expression of these proteins and disease prognosis.. In the sample, the presence of immunohistochemically detectable p53 is associated with shorter survival and disease-free intervals, as shown in Kaplan-Meier survival curve analysis. Indeed, multivariate statistical analysis revealed that the accumulation of p53 is an independent prognostic factor, which is associated with shorter survival. This association was not evident in the case of cyclin D1.. A more precise prognosis of patients with laryngeal epidermoid carcinomas could be achieved by taking into account the presence of p53 (as assayed by immunohistochemical analysis) in biopsy tissue

Topics: Adult; Aged; Biomarkers, Tumor; Biopsy, Needle; Carcinoma, Squamous Cell; Cyclin D1; Female; Genes, p53; Genetic Predisposition to Disease; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Multivariate Analysis; Mutation; Neoplasm Staging; Prognosis; Proportional Hazards Models; Reference Values; Retrospective Studies; Sensitivity and Specificity; Survival Analysis

Alterations of cell cycle proteins contribute to the development and biological behaviour of malignant tumours. We evaluated the distribution and prognostic significance of immunohistochemically detected proteins p53, p21, Rb, and cyclin D1 in 101 laryngeal and hypopharyngeal squamous cell carcinomas (SCC) and adjacent epithelial hyperplastic lesions (EHL). Protein expression was correlated with tumour grade and stage. Varying patterns of protein expression were found in SCC. A significant correlation (p<0.05) was found between Rb expression and tumour grade. Different grades of EHL exhibited randomly distributed p53 and cyclin D1 positive cell clusters with no association to the pattern of their expression in SCC. Our study demonstrated derailment of cell cycle regulation in almost all cases of SCC of the larynx and hypopharynx. However, only cyclin D1 expression had an independent prognostic value for cancer-specific survival. The results also suggest that Rb gene inactivation, although rare, might be more important in the development of SCC than previously thought.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Enzyme Inhibitors; Female; Humans; Hyperplasia; Hypopharyngeal Neoplasms; Hypopharynx; Immunohistochemistry; Laryngeal Neoplasms; Larynx; Male; Middle Aged; Prognosis; Respiratory Mucosa; Retinoblastoma Protein; Tumor Suppressor Protein p53

The prognosis in patients suffering from head and neck squamous cell carcinomas depends on many factors. However, regional lymph node metastases are the most important parameter in determining the cure and survival of patients with head and neck cancers. The evaluation of cancer cell biology enables differentiation of their proliferation and tendency of metastases. Immunohistochemical examinations complement the well-established routine histological examination. The aim of this study was to evaluate the prognostic importance of the level of immunoproliferating proteins such as cyclin D1, nuclear antigen Ki67 and suppressor gene p53 for regional lymph node metastases in laryngeal carcinoma. The research was carried out on 73 patients treated for squamous cancer of the larynx in the Department of Otolaryngology University School of Medical Sciences in Poznan in the years 1994-1999. The group was comprised of 4 female and 69 male patients. Their ages ranged from 37 to 79 years, with a mean of 59 years. Clinical data included sex, age, localization and local and regional extent of the tumor, presence or lack of distant metastases, treatment, histological examination as well as immunohistochemical evaluation of suppressor gene p53, proliferative proteins Ki67 and cyclin D1. No statistically significant correlation was found between staining intensity of suppressor gene p53, cyclin D1 and the degree of local advancement (T). There was no correlation between the level of immunoproliferative markers and regional lymph node metastases. Statistically significant correlation was found between T stage and staining for Ki67 (P=0.017) as well as between cyclin D1 level and Ki67 (P<0.05). In conclusion, (1) no significant correlation was found between Ki67 and cyclin D1, p53 and TNM classification; (2) lack of correlation was confirmed between N+, p53, Ki67, cyclin D1 and Jacobsson classification; (3) the degree of histological grading correlated, however, with Jacobsson classification and cyclin D1 expression.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma; Cyclin D1; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neck; Prognosis; Tumor Suppressor Protein p53

At present, the differences between head and neck basaloid squamous cell carcinoma (BSCC) and nonbasaloid squamous cell carcinoma (SCC) are mostly on the basis of histologic and immunohistologic findings.. In this study, we investigated 8 BSCCs and 59 SCCs for loss of heterozygosity (LOH) at chromosomes 5q, 9p, 9q, 10q, 11q, 13p, 17p, and 18q. In addition, we analyzed p16, PTEN, and CCND1 (cyclin D1) and investigated the HPV status. Immunohistochemically, the expression of MIB-1, p16, p53, and cyclin D1 was determined.. Aberrations in the BSCCs were especially frequent at 9p and in the CCND1 gene. In contrast, alterations at 10q occurred almost exclusively in conventional SCCs. Obvious differences could be determined concerning the HPV status: HPV-DNA was detected in all BSCCs but only in 17% of conventional SCCs.. Although the number of investigated BSCCs is rather low and did not allow statistical conclusions, our results focus on certain differences between the molecular pathogenesis of BSCCs and SCCs.

Topics: Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA, Viral; Genes, cdc; Head and Neck Neoplasms; Humans; Immunohistochemistry; Laryngeal Neoplasms; Loss of Heterozygosity; Microsatellite Repeats; Papillomaviridae; Phosphoric Monoester Hydrolases; PTEN Phosphohydrolase; Tumor Suppressor Proteins

The recent development of tissue array technology has potentiated large-scale retrospective cohort studies using archival formalin-fixed, paraffin-embedded tissues. This study evaluates the potential for using archival head and neck cancer tissue in such arrays.. Tissue array blocks were made from 184 head and neck cancer specimens. Three core tissue biopsies (0.6 mm x 3-4 mm) were taken from individual "donor" paraffin-embedded tumor blocks and arrayed into a new "recipient" paraffin block. Immunohistochemical (IHC) analyses were performed using antibodies recognizing cyclin-D1, Rb, and EGFR. IHC was scored on a 6-point scale for extent and a 3-point scale for intensity. We compared the staining of tissue array disks with staining of full tissue sections.. Seventy-four percent (475 of 640) of samples placed into tissue arrays were confirmed to represent tumor tissue. The remaining samples were lost during processing or contained too few tumor cells. Only 6% of cases were completely lost, whereas 55%, 28%, and 11% of cases were judged on 3, 2, or 1 disk, respectively. Cohen's kappa coefficient was 0.66 for cyclin-D1, 0.40 for EGFR, and 0.41 for Rb.. Tissue array technology is a rapid and efficient method for retrospective analysis of protein expression and is a promising tool for validation of prognostic markers in large series of head and neck squamous cell carcinomas. The agreement in scoring of the full section and the tissue arrays is reasonable. Discordance is probably due to intraobserver variation and lack of robustness of the scoring inherent of the proteins studied.

Topics: Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; ErbB Receptors; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Oropharyngeal Neoplasms; Prognosis; Protein Array Analysis; Retinoblastoma Protein; Statistics, Nonparametric

In order to investigate the telomerase activity and its relationship with the overexpression of cyclin D1 protein in laryngeal squamous cell carcinomas (LSCC).. TRAP(Telomeric repeat amplification protocol) assay and immunohistochemical method were used respectively to detect the telomerase activity and the overexpression of cyclin D1 protein in 38 LSCC and correspondent laryngeal mucosa.. The expressions of telomerase and cyclin D1 protein were 82%(31/38), 50%(19/38) respectively in LSCC, both of them were not associated with the age of patients, location of tumors, T stage and pathological grade(P > 0.05), but there was a significant difference among clnic stages (P < 0.05) in telomerase activity, no significant difference could be found among clinic stages in expression of cyclin D1. 18 out of 31 cases were expressed in both of them simultaneously there was significant association between them(P < 0.05).. Telomerase is widely activated in LSCC, the overexpression of cyclin D1 protein may be one of the most important mechanisms of telomerase activation in LSCC.

Topics: Carcinoma, Squamous Cell; Cyclin D1; Humans; Laryngeal Neoplasms; Telomerase

To find out whether the expression of p27 and cyclin D1 is different in laryngeal squamous cell carcinoma and the edge tissues, and to study the relationship between the expression of p27 and cyclin D1 with the clinic character in patients with laryngeal squamous cell carcinoma.. An immunohistochemistry method was used to study the expression of p27 protein and cyclinD1 in 38 laryngeal squamous cell carcinoma and the edge tissues.. The expression of p27 protein and cyclin D1 was significantly different between the laryngeal carcinoma and the edge tissue. Low expression of p27 and over expression of cyclinD1 was associated with the liability of lymph node metastasis and the poor prognosis for patient with laryngeal squamous cell carcinoma. An interesting observation was that the inverse expression between p27 and cyclinD1 might play an important role in laryngeal squamous cell carcinoma.. p27 protein and cyclinD1 in laryngeal carcinoma is apparently a significance predictor of survival.

Topics: Aged; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Microfilament Proteins; Middle Aged; Muscle Proteins; Prognosis

Cyclin D1 is a regulatory factor essential in the progression of the cell cycle from G1 through S phase. Amplification and overexpression of cyclin D1 have been observed in many human cancers including head and neck squamous cell carcinoma (HNSCC). We have previously transfected a HNSCC control cell line (CCL23) with an antisense cyclin D1 plasmid and demonstrated inhibition of cell proliferation in vitro. In this study, we examine whether antisense cyclin D1 could inhibit tumor growth in vivo. Methods/measures: The CCL23 and its antisense cyclin D1 transfected clone (CCL23 AS) were injected into the flanks of nude mice. Tumor growth was monitored weekly. After 5 weeks, tumors were removed and studied for tumor size, cyclin D1 expression, cyclin D1-dependent kinase activity, and retinoblastoma (Rb) phosphorylation.. Compared with the control tumors, 11 of 19 antisense tumors were smaller, 7 tumors were of equal size, and 1 tumor was larger. Immunohistochemical analysis with an anti-cyclin D1 antibody demonstrated decreased cyclin D1 expression in CCL23 AS and the smaller antisense tumors. Cyclin D1-dependent kinase activity was reduced in CCL23 AS and the smaller antisense tumors, and this was accompanied by a relative decrease in phosphorylated Rb in these samples.. Antisense cyclin D1 inhibits growth of HNSCC tumors. Cyclin D1 expression, cyclin D1-dependent kinase activity, and Rb phosphorylation are decreased in these tumors.. These findings lend support for the potential use of antisense cyclin D1 as gene therapy for HNSCC.

Topics: Animals; Antisense Elements (Genetics); Blotting, Western; Carcinoma, Squamous Cell; Cyclin D1; Female; Immunohistochemistry; Laryngeal Neoplasms; Mice; Mice, Nude; Phosphorylation; Transfection; Transplantation, Heterologous; Tumor Cells, Cultured

Cyclin D1 and its catalytic partner CDK4 are known to play important roles in the G1/S checkpoint of the cell cycle. The complex formed by CDK4 and cyclin D1 has been strongly implicated in the control of cell proliferation and prognoses in human malignancies. We investigated the immunohistochemical expression of cyclin D1, CDK4 and proliferating cell nuclear antigen (PCNA) in 102 patients with laryngeal squamous cell carcinoma (LSCC). Cyclin D1 overexpression was observed in 59 cases (57.8%) of LSCC, and was significantly correlated with tumor site, tumor size, lymph node metastasis and advanced stage. CDK4 overexpression was observed in 48 cases (47.1%), and was significantly correlated with tumor size and advanced stage. Cyclin D1 and CDK4 expression was significantly associated with cell proliferation measured by PCNA (r = 0.812, p < 0.0001 and r = 0.725, p < 0.0001, respectively). The Kaplan-Meier analysis showed that cyclin D1 overexpression was significantly associated with disease-free survival and overall survival. CDK4 overexpression was significantly associated with overall survival. When cyclin D1 and CDK4 are combined, the patients with co-overexpression of cyclin D1-CDK4 revealed the poorest overall survival. Additionally, in early-stage (I-II) cases, co-overexpression of cyclin D1-CDK4 was also revealed to possess a significant prognostic role. By multivariate analysis, cyclin D1 overexpression, lymph node metastasis and advanced stage were independent prognostic factors for disease-free survival. Cyclin D1 overexpression, CDK4 overexpression, tumor grade, lymph node metastasis and advanced stage were independent prognostic factors for overall survival. These findings indicate that cyclin D1 and CDK4 overexpression and/or co-overexpression of these proteins may play a pivotal role in the biological behavior of LSCC and may provide a strong prognostic implication.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; China; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinases; Disease-Free Survival; Female; Follow-Up Studies; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Multivariate Analysis; Prognosis; Proto-Oncogene Proteins; Regression Analysis; Survival Rate

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

Recent laboratory experiments have demonstrated that cyclin D1 levels (cycD1) can influence radiosensitivity. The purpose of the current study is to evaluate the prognostic significance of cycD1 for local recurrence in early-stage larynx cancer treated with primary radiation therapy. The study was conducted using a matched case-control design in 60 early-stage (T1-T2/N0) larynx cancer patients. All patients had squamous cell carcinoma of the larynx and were treated with primary radiation to a total median dose of 66 Gy in daily fractions of 2 Gy, without surgery or chemotherapy. Thirty patients who suffered a local relapse in the larynx after treatment served as the index case population. These 30 cases were matched by age, sex, site (glottic vs. supraglottic), radiation therapy technique/dose, and follow-up, to 30 control patients who did not experience a local relapse. Immunohistochemical staining from cycD1 was performed on the paraffin-embedded specimens. The pathologist, blinded to the clinical information, scored each of the specimens on a four-point intensity scale (0 = no stain, 1 = faint, 2 = moderate, 3 = strong) and percent distribution. Patients were considered to be positive for cyclin D1 if the staining was 2+ or greater with a percent distribution of at least 5%. By design of the study, the two groups were evenly balanced with respect to age, sex, stage, radiation dose, and follow-up. CycD1 levels correlated with proliferating cell nuclear antigen levels. Low levels of cycD1 significantly correlated with local relapse; 19/30 (63%) of the index cases stained negative, while only 10/30 (33%) of the control cases stained negative (P = 0.03). These data suggest that low levels of cycD1 correlate with relatively radioresistant early-stage larynx carcinoma. With larger more confirmatory clinical and laboratory data, this data may have significant clinical implications. Int. J. Cancer (Radiat. Oncol. Invest.) 90, 22-28 (2000).

Topics: Carcinoma, Squamous Cell; Case-Control Studies; Cyclin D1; Female; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; Proliferating Cell Nuclear Antigen; Radiotherapy Dosage

To study the correlation between the expression level of Cyclin D1 and the laryngeal squamous cell carcinoma (LSCC).. 35 LSCC cases testified pathologically were included in the study. The samples obtained from each tumor tissue and adjacent normal mucosa were detected to see the expression level of Cyclin D1 with the method of RT-PCR and light density half quantive analysis.. The expression level of Cyclin D1 was significant higher in almost all turmor tissues than in the adjacent normal mucosae (P < 0.05). Overexpressed carcinomas were more frequently associated with lymph node metastases (P < 0.05). A significant association between the expression level of Cyclin D1 and TNM was found. The higher T stage, the higher the expression level of Cyclin D1 (P < 0.01). The expression level of low histological grade carcinomas was lower than that of the high histological grade carcinomas, but the difference had no statistical significance. No association between overexpression of Cyclin D1 and sex, age was found too.. The result indicates that the expression level of Cyclin D1 in LSCC may be a useful marker of prognostic significance and it provide a molecular biology basis for treating the LSCC patients.

Topics: Aged; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Prognosis; Reverse Transcriptase Polymerase Chain Reaction

Cyclin D1, p16, and Rb genes play a critical role in the regulation of the G1-S transition of the cell cycle and are frequently altered in several neoplastic entities. Analysis of the protein products of these genes by molecular and immunohistochemical methods provides information on their functional status and allows for the phenotypic evaluation of tumor cells. We performed Western blotting and immunohistochemical analysis on tissues from 35 primary oral and laryngeal squamous carcinoma specimens with previous molecular analysis of the p16 gene and correlated the results with relevant clinicopathologic factors. Our study shows significant concordance between Western blotting and immunostaining results for cyclin D1 (P = .01), p16 proteins (P = .01), and Rb (P = .04). Heterogeneous staining of tumor cells and the positivity of non-neoplastic host elements for Rb by immunohistochemistry contributed to the discrepancy noted in some tumors by Western blotting. Significant reciprocal relationship between p16 and Rb proteins was observed (P < .001); in most tumors, absence of p16 (89%) and detectable Rb (94%) proteins were found. Two tumors had negative cyclin D1 expression, and one third overexpressed this protein. There was a lack of correlation between cyclin D1 overexpression and the clinicopathologic factors studied. Our results indicate that the absence of p16 in most of these tumors may constitute an early tumorigenic event and that the loss of the Rb function plays a minor role in HNSC.

Topics: Adult; Aged; Aged, 80 and over; Blotting, Western; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA Methylation; Female; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Mutation; Retinoblastoma Protein

GST, CYP, and CCND1 genotypes have been associated with outcome in several cancers. Accordingly, we have examined, in patients with one squamous cell carcinoma (SCC) of the head and neck, associations between GSTM1, GSTT1, GSTM3, GSTP1, CYP2D6, CYP1A1, CYP2E1, and CCND1 genotypes and the outcome parameters, tumor extension, histological grade, and presence of nodes. We used logistic regression to study, first, each gene individually and, second, in a step-wise model that included all of the genes. Different genes were associated with each outcome parameter. Thus, GSTT1 null was associated with T3/T4 lesions in the oral cavity/pharyngeal (P = 0.029), but not laryngeal, SCC cases. GSTT1 null was also associated with histological differentiation (G3) in the oral cavity/pharyngeal, but not laryngeal, SCC cases, although this association only approached significance (P = 0.069). CCND1 GG was associated with G3 tumors in the oral cavity/pharyngeal (P = 0.011), but not laryngeal, SCC cases. The combination of GSTT1 null/CCND1 GG was also associated with G3 tumors. CYP2D6 PM and HET were associated with lymph node involvement in the laryngeal, but not oral/pharynx, SCC cases. Genes that were individually associated with outcome were also associated with the parameter in the step-wise routine. The GSTT1 null frequency was greater in 39 patients with second primary tumors than in those with one lesion (P = 0.014). The data demonstrate site-dependent associations between GSTT1 null, CCND1 GG, and CYP2D6 PM and tumor extension, differentiation, and nodes.

Topics: Carcinoma, Squamous Cell; Cyclin D1; Cytochrome P-450 Enzyme System; Female; Genotype; Germany; Glutathione Transferase; Humans; Laryngeal Neoplasms; Male; Middle Aged; Multivariate Analysis; Oropharyngeal Neoplasms; Polymerase Chain Reaction

Overexpression of cyclin D1 has been found in a variety of malignancies and is suggested to be related to tumor progression. We immunohistochemically investigated the overexpression of cyclin D1 protein in 92 laryngeal carcinomas. Twenty-eight (30.4%) of the carcinoma specimens showed overexpression of cyclin D1. This overexpression was not related to the tumor stage, lymph node metastasis, or clinical outcome. However, the overexpression of cyclin D1 in patients with local recurrence was significantly higher than in patients with no recurrence. Cyclin D1 immunohistochemical staining is considered to be a useful marker for predicting tumor recurrence.

Topics: Biomarkers, Tumor; Cyclin D1; Female; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Neoplasm Recurrence, Local

Expression of interrelated gene products regulating cell proliferation and apoptosis may be disordered in squamous cell carcinoma (SCC) of the larynx compared with normal squamous mucosa. Certain of these abnormalities, alone or in combination, may be of prognostic significance in low-stage carcinomas of the larynx. A retrospective study of archival material was made. Expression of the Bcl-2 family of apoptosis-related genes (bcl-2, bcl-X, mcl-1, and bax) and the proliferation- and apoptosis-related genes p53 and cyclin D-1 were determined in 40 low-T-stage laryngeal carcinomas and in uvular epithelium from patients without SCC. Among the antiapoptotic members of the Bcl-2 family, Bcl-X and Mcl-1 showed more intense and widespread staining than Bcl-2 itself in both normal squamous mucosa and SCC. The well-ordered expression patterns of Bcl-2-related proteins found in normal epithelium were lost in SCC, and patterns of expression varied widely among individual tumors. Also, mean expression levels for Bax and cyclin D-1 were significantly lower than in normal epithelium (P = .036 and P = .009, respectively), whereas expression of p53 was higher in tumors (P = .034). Expression of Bcl-X and Mcl-1 was greater in poorly differentiated than in well-differentiated tumors (P = .014 and P = .031, respectively). No associations were seen between marker expression patterns and clinical outcome in this group of patients. Bcl-x and Mcl-1 appear to be the most abundantly expressed antiapoptotic proteins of the Bcl-2 family in both normal squamous mucosa and SCC of the larynx. Multiple genes regulating proliferation and apoptosis are expressed abnormally in laryngeal SCC compared with normal epithelium. In particular, loss or measurable decrease in expression of the proapoptotic protein Bax in tumors may contribute to the deranged growth control of SCC. Further study is needed to evaluate the prognostic significance of particular patterns of disordered expression of proteins regulating proliferation and apoptosis in SCC of different head and neck sites.

Topics: Adult; Aged; Aged, 80 and over; Apoptosis; Carcinoma, Squamous Cell; Cell Division; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Genes, bcl-1; Genes, bcl-2; Genes, p53; Humans; Laryngeal Neoplasms; Male; Middle Aged; Mucous Membrane; Proto-Oncogene Proteins c-bcl-2; Retrospective Studies; Tumor Suppressor Protein p53

To investigate the prognostic relevance of cyclin D1 gene overexpression in laryngeal squamous cell carcinomas (LSCCs).. The overexpression of cyclin D1 was analyzed in 149 LSCC patients with a median follow-up duration of 60 months using the DCS6 monoclonal antibody; only cases that overexpressed cyclin D1 in more than 5% of neoplastic cells were considered positive.. Forty-eight cases (32.2%) were immunoreactive to the DCS6 antibody. Cyclin D1 overexpression was significantly associated with tobacco smoking and alcohol consumption, tumor extension, advanced clinical stage, and the presence of lymph node metastases. Univariate analysis showed that a shorter disease-free and overall survival were significantly associated with supraglottic site, tumor extension, advanced clinical stage, and cyclin D1 overexpression. At multivariate analysis, tumor extension and cyclin D1 overexpression were significantly associated with tumor recurrence, whereas tumor extension, supraglottic site and, at a borderline level of statistical significance, cyclin D1 overexpression, were associated with reduced overall survival.. The overexpression of cyclin D1 in LSCC is associated with unfavorable clinicopathologic features and represents an independent significant predictor of laryngeal carcinoma prognosis, particularly for disease-free survival. This indicates that cyclin D1 evaluation may be a further useful element for selecting subgroups of patients who should be treated with more aggressive therapies.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; Disease-Free Survival; Female; Follow-Up Studies; Gene Expression; Humans; Immunohistochemistry; Laryngeal Neoplasms; Male; Middle Aged; Prognosis; Survival Analysis

To evaluate the predictive role of the oncogenes p53, MDM-2 and cyclin D1, and the proliferative marker Ki67, in the progression from low-grade dysplasia to carcinoma of the larynx. We studied immunohistochemically a series of 32 low-grade pre-neoplastic laryngeal lesions, 10 of which progressed to invasive carcinoma. Immunoreactivity in more than 10 per cent of the dysplastic cells was detected in five cases immunostained with anti-p53 (approximately 15 per cent), in two with anti-MDM-2 (approximately six per cent), and 11 with anti-Ki67 antibodies (approximately 34 per cent), whereas none of the cases showed cyclin D1 overexpression. No significant association was found between p53 and MDM-2 immunoreactivity and the evolution to carcinoma; on the contrary, Ki67 expression was detectable in all but one of the 10 cases developing an infiltrative tumour (90 per cent), and in two of the 22 cases that did not progress (approximately nine per cent) (p = 0.01). These findings indicate that immunohistochemical assessment of the proliferative index in bioptic samples of dysplastic laryngeal mucosa may be useful in selecting patients who should undergo a more specific follow-up evaluation.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cyclin D1; Disease Progression; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Ki-67 Antigen; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Proteins; Nuclear Proteins; Precancerous Conditions; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-mdm2; Tumor Suppressor Protein p53

Head and neck squamous cell carcinomas (SCCs) seem to follow a multistep process of carcinogenesis in which chemical and/or viral agents are associated with specific genetic alterations. The prevalence of human papillomavirus (HPV) infection and the amplification of the cyclin D1 (CCND1) gene were evaluated in a series of 75 laryngeal SCCs by PCR with HPV consensus primers and Southern blot analysis with a CCND1-specific probe, respectively. HPV DNA was detected in 22 of 75 (29.3%) tumors, and it belonged almost exclusively to the highly oncogenic HPV-16, HPV-18, and HPV-33. CCND1 gene amplification was found in 15 of 75 (20%) tumors, and it was associated with HPV infection in a statistically significant manner (chi2 = 20.3; P < 0.001). Because the viral oncoproteins E6 and E7 from high-risk HPV types are known to promote genomic rearrangements, these findings suggest that amplification of the CCND1 gene in laryngeal SCCs may occur as a consequence of the genomic instability associated with HPV infection. In turn, amplified CCND1, either alone or in conjunction with a direct action of the viral oncoproteins E6 and E7, could lead to a perturbation of the cell cycle. This model could explain the involvement of high-risk HPV types in laryngeal carcinogenesis.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; DNA, Viral; Female; Gene Amplification; Humans; Laryngeal Neoplasms; Male; Middle Aged; Papillomaviridae; Papillomavirus Infections; Tumor Virus Infections

The molecular pathogenesis of laryngeal squamous cell carcinomas (LSCCs) is still only partially understood, although genetic alterations affecting various protooncogenes or tumor suppressor genes have often been detected.. To improve their understanding of the role of cyclin D1 in the pathogenesis of LSCCs, the authors investigated the expression of cyclin D1 protein and the amplification status of the bcl-1/cyclin D1 locus in a panel of 58 pathologic samples.. Expression of cyclin D1 protein was detected in 23 of the 58 patients (approximately 39%), 14 of whom had lymph node metastases (approximately 61%); of the remaining 35 patients without any detectable cyclin D1 expression, 7 had lymph node metastases (20%). Expression of cyclin D1 was detectable in 5% of the specimens of normal mucosa, 13% of those with mild-to-moderate dysplasia, and 25% of those with severe dysplasia. Amplification of the bcl-1/cyclin D1 locus was detected in 12 of the 49 LSCCs investigated (approximately 24%), 7 of which had lymph node metastases (approximately 58%); of the remaining 37 LSCCs with an apparently normal copy number of the cyclin D1 locus, 12 had lymph node metastases (approximately 32%). The authors found almost complete concordance between locus amplification and protein expression. Statistical analysis showed a correlation between cyclin D1 expression and both the presence of lymph node metastases (P < 0.01) and advanced clinical stage (P < 0.02).. The authors' observations suggest that the deregulation of cyclin D1 expression may be involved in the pathogenesis of more aggressive LSCCs.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; Cyclins; Female; Gene Amplification; Gene Expression Regulation, Neoplastic; Genes, bcl-2; Humans; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Oncogene Proteins

Topics: Antibodies, Neoplasm; Biomarkers, Tumor; Biopsy; Carcinoma, Squamous Cell; Cell Division; Cyclin D1; Cyclins; Humans; Immunohistochemistry; Ki-67 Antigen; Laryngeal Neoplasms; Nuclear Proteins; Oncogene Proteins; Predictive Value of Tests; Prognosis; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-mdm2; Tumor Suppressor Protein p53

Squamous carcinoma of the larynx arises from pre-existing lesions, the so-called "preneoplastic lesions". Hyperplastic lesions represent a part of their spectrum, from both clinical and biological points of view. On morphologic grounds, the most characteristic feature with prognostic value in the evaluation of preneoplastic lesions is dysplasia. It is not only nuclear alterations that are seen in the process of malignant transformation, the cytoplasmic pattern of cytokeratins changes through neoplastic progression, with a progressive reduction of the molecular weight of the produced species. Dysplasia also associates with gross alterations of the DNA content. This is in agreement with our finding of alterations of genes participating in the control of the cell cycle, p53 and p21(WAF1/cip1). p53 overexpression is detected in non-invasive squamous lesions (even in the absence of obvious dysplasia) and p21(WAF1/cip1) shows a dramatic change in the pattern of expression in dysplastic epithelium compared with the normal. However, not all genes participating in the control of the cell cycle are altered in early lesions. Overexpression of cyclin D1, a common phenomenon in advanced carcinomas, is not likely to participate in the early phases of neoplastic development.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Cyclin D1; Cyclin-Dependent Kinases; Cyclins; DNA, Neoplasm; Epithelium; Genes, p53; Humans; Hyperplasia; Keratins; Laryngeal Diseases; Laryngeal Mucosa; Laryngeal Neoplasms; Oncogene Proteins; Precancerous Conditions

The expression of cyclin D1 gene was investigated in 74 laryngeal squamous cell carcinomas (LSCCs) in order to determine its clinical and prognostic value. Overexpression of cyclin D1 was detected immunohistochemically using DCS6 monoclonal antibody on formalin-fixed, paraffin-embedded tissue sections. Cyclin D1 expression was detected in 22 of the 74 cases investigated (30 per cent), thirteen of which presented nodal metastases (59 per cent); of the patients without any detectable cyclin D1 protein expression, six presented nodal metastases (12 per cent). Cyclin D1 protein expression was found in five per cent of the specimens of normal mucosa, eight per cent of those with low-grade dysplasia and 20 per cent of those with high-grade dysplasia. A statistically significant association was found between cyclin D1 expression and the supraglottic site (p < 0.05), tumour extension (p < 0.001), the presence of lymph node metastases (p < 0.001), and advanced clinical stage (p < 0.001). Cyclin D1 expression analysis is an important tool in the selection of LSCC patients with an aggressive clinical course.

Topics: Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; Cyclins; Disease Progression; Female; Humans; Immunohistochemistry; Laryngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Oncogene Proteins

We have examined the presence of p16MTS1/CDK4I gene deletions, mutations and methylation status, and 9p21-23 deletions in a series of 46 squamous cell carcinomas of the larynx and paired normal mucosa previously characterized for cyclin D1 gene amplification and overexpression. pRb expression was also examined by immunohistochemistry. p16MTS1/CDK4I mutations were found in 10/46 (22%) carcinomas and hypermethylation in 2/31 (7%). Loss of heterozygosity at 9p21-23 was found in 24 out of 42 (57%) carcinomas examined. All p16MTS1/CDK4I mutated cases and the two hypermethylated carcinomas showed 9p21-23 loss of heterozygosity. The loss of heterozygosity correlated with advanced local invasion (P=0.0045), lymph node metastases (P=0.0326), stage IV of the tumors (P=0.0058), and existence of cyclin D1 amplification/overexpression (P < 0.03). Only one out of 37 carcinomas was negative for pRb expression. No alterations in p16 gene or 9p21-23 loss of heterozygosity were detected in this case. These findings indicate that p16MTS1/CDK4I is frequently inactivated by gene mutation, hypermethylation, and allelic deletions in a significant subset of squamous cell carcinomas of larynx. Since 9p21-23 loss of heterozygosity was more frequently detected than p16MTS1/CDK4I mutations, and mutated carcinomas invariably had loss of heterozygosity, allelic losses probably precede the p16MTS1/CDK4I mutations. Their association with cyclin D1 deregulation in advanced carcinomas could indicate a possible cooperative effect in the progression of these neoplasms.

Topics: Aged; Amino Acid Substitution; Carcinoma, Squamous Cell; Chromosome Mapping; Chromosomes, Human, Pair 9; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Frameshift Mutation; Humans; Laryngeal Neoplasms; Loss of Heterozygosity; Lymphatic Metastasis; Male; Microsatellite Repeats; Middle Aged; Mutation; Neoplasm Invasiveness; Neoplasm Staging; Point Mutation; Polymerase Chain Reaction; RNA Splicing; Sequence Deletion

To determine the effect of gene of carcinorma on the proliferation and regulation in primary laryngeal cancer, P16, Rb, Cyclin D1 were examined with immunohistochemical SABC method in 36 cases of primary laryngeal cancer. The results showed: P16 was positive in 11 of 36 in carcinoma specimens (30.6%), 22 of 36 in the specimens surrounding the carcinoma (61.1%) were positive. The difference was significant (P < 0.05). The positive rates of Rb and Cyclin D1 were 61.1% and 47.2% respectively. Comparing the positive rate of P16 to Rb was more significantly (P < 0.05). In most circumstances, when Rb was positive, P16 was negative and vice versa. The positive rate of Rb correlated with the one of Cyclin D1 positively by rank correlation. The expression of P16 and Rb was significantly different between well and poor tissues differenciation. The expression had no significant difference in the same antibody in various clinical stage and position or in the same stage and position but different antibody. The results suggest that the feedback regulation chain consisted of P16 plays an important role in the proliferation and regulation.

Topics: Aged; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Gene Expression; Genes, bcl-1; Genes, p16; Genes, Retinoblastoma; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Staging; Retinoblastoma Protein

The cyclin D1 (CCND1) gene is amplified, rearranged, and overexpressed frequently in human cancer, including squamous cell carcinoma. The gene dosage of CCND1 was examined in 51 primary laryngeal squamous cell carcinomas, and amplification of the gene was found in 9 (17.6%) cases. CCND1 amplification did not correlate with age, tumor localization and extension, cervical lymph node involvement, histopathological grading, and epidermal growth factor receptor levels. In a univariate analysis, CCND1 amplification, tumor extension, lymph node involvement, poor histological differentiation, and high epidermal growth factor receptor levels were correlated significantly with shorter overall survival. In a median follow-up period of 29 months, the overall survival rate was 71.4% for patients affected with tumors displaying a normal CCND1 dosage and only 25.0% for patients affected with tumors carrying amplified CCND1 (P = 0.0288). In a multivariate analysis, only CCND1 and tumor extension retained statistically significant prognostic values (P = 0.037 and 0.041, respectively). This is the first report in which CCND1 amplification is identified as a significant independent prognostic factor in laryngeal carcinoma. Evaluation of CCND1 amplification could be applicable to the clinical management of laryngeal cancer, allowing identification of patients with poor prognoses.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; ErbB Receptors; Female; Gene Amplification; Humans; Laryngeal Neoplasms; Male; Middle Aged; Prognosis; Survival Rate

To understand the molecular pathogenesis of laryngeal squamous cell carcinomas (LSCCs), this study investigated the involvement of various protooncogene loci (bcl-1, int-2, c-erbB-1, c-myc, ras) and the p53 tumor suppressor gene in 18 patients with LSCC (15 at clinical presentation, 3 in clinical relapse).. For all patients, the mutations affecting the p53 and the H-, K-, and N-ras genes were evaluated by polymerase chain reaction (PCR), single-strand conformation polymorphism, and the direct sequencing of PCR-amplified fragments. The bcl-1, int-2, c-erbB-1, and c-myc loci of 15 patients were investigated using Southern blot analysis.. A mutation of the p53 gene was detected in 5/18 patients (approximately 28%), bcl-1 locus amplification in 4/15 (approximately 26%), c-erbB-1 locus amplification in 2/15 (approximately 13%), and c-myc locus amplification in 1/15 (approximately 6%). The simultaneous presence of more than one genetic lesion was observed in four patients; two showed int-2/bcl-1 coamplification, and two int-2/c-erbB-1 coamplification, one of whom also showed a p53 gene mutation. A novel p53 mutation involving the splice acceptor site of exon 6 was detected in one patient. Two of the five patients positive for p53 mutations had clinical relapses of primary tumors. bcl-1 locus amplification only was observed in patients with lymph node metastases (4/6). All but one of the patients with molecular genetic lesions showed a peculiar infiltrating pattern.. Overall, these results show that alterations of known protooncogenes and the p53 tumor suppressor gene are involved in a large proportion of LSCCs (11/18; approximately 60%) and may suggest that distinct molecular pathways occur in the pathogenesis of these tumors.

Topics: Aged; Base Sequence; Blotting, Southern; Carcinoma, Squamous Cell; Cyclin D1; DNA Mutational Analysis; DNA, Neoplasm; Fibroblast Growth Factor 3; Fibroblast Growth Factors; Genes, erbB; Genes, myc; Genes, p53; Genes, ras; Humans; Laryngeal Neoplasms; Male; Middle Aged; Molecular Sequence Data; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Proto-Oncogene Proteins; Proto-Oncogenes

PRAD-1 is a putative oncogene localized on chromosome 11q13 which encodes cyclin D1, a novel cyclin involved in cell cycle regulation. Amplification of this gene has recently been reported in several human tumors including breast and head and neck carcinomas. In this study we have analyzed the presence of PRAD-1/cyclin D1 gene amplification and mRNA overexpression in a series of 46 matched normal mucosas and squamous cell carcinomas of the larynx. PRAD-1/cyclin D1 was found to be amplified 2- to 12-fold in 17 carcinomas (37%). DNA amplification correlated with advanced local invasion (P = 0.0015), presence of lymph node metastases (P = 0.0078), and stage IV of the tumors (P = 0.0021). mRNA overexpression was found in 15 of the 43 (35%) cases examined and it was also significantly associated with advanced local invasion (P = 0.0025) and stage IV carcinomas (P = 0.0032). A significant association was observed between gene amplification and mRNA overexpression (P < 0.0001) with only 3 discordant cases (2 amplifications with no overexpression and 1 overexpressed carcinoma with no gene amplification). Furthermore, the degree of DNA amplification correlated with the levels of mRNA expression (r = 0.6; P = 0.024). These findings suggest that the PRAD-1/cyclin D1 gene may be an important target of 11q13 amplifications in laryngeal carcinomas and the activation of this gene may be involved in the progression of these tumors. Its association with advanced-stage tumors indicates that PRAD-1/cyclin D1 gene amplification and overexpression may be of prognostic significance.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Chromosomes, Human, Pair 11; Cyclin D1; Cyclins; DNA, Neoplasm; Gene Amplification; Humans; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Oncogene Proteins; RNA, Messenger; RNA, Neoplasm