Distribution and prognostic significance of cell cycle proteins in squamous carcinoma of the larynx,hypopharynx and adjacent epithelial hyperplastic lesions

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.     

Expresión de E-cadherina y β-catenina en carcinomas de células escamosas de laringe e hipofaringe

Introduction and objectivesDysfunction of the E-cadherin/catenin complex is directly related to carcinogenesis and metastases development.The aim of this paper is to investigate the prognostic significance of E-cadherin and β-catenin expression in surgically treated laryngeal and hypopharyngeal squamous cell carcinomas.Material and methodsTumour tissue samples were obtained from 133 consecutive patients with squamous cell carcinomas of the head and neck: 68 of the larynx and 65 hypopharyngeal carcinomas, who underwent surgical treatment in our hospital between 2000 and 2005.E-cadherin and β-catenin expression was analysed by immunohistochemistry, quantifying the percentage of stained cells and the intensity of staining.ResultsE-cadherin and β-catenin expression was evaluable in 59 laryngeal carcinomas and in 58 cases of hypopharyngeal carcinomas.In the laryngeal tumours, a significant association was found between the low expression of membrane β-catenin with T4 tumours and tumour recurrence. In the hypopharynx there was a significant association between positive expression of nuclear β-catenin and poor histological differentiation (P = .02). In the multivariate analysis, only the presence of lymph node metastases was an independent predictive factor of decreased disease-specific survival in laryngeal squamous cell carcinomas.ConclusionsThe expression of E-cadherin and β-catenin does not show prognostic significance in laryngeal and hypopharyngeal squamous cell carcinomas over the TNM classification.     

Prognostic value of the expression of p53 and bcl-2 in patients with laryngeal carcinoma

Previous studies have yielded inconsistent results on the prognostic significance of p53 and bcl-2 in head and neck cancer. The aim of this study was the evaluation of p53 and bcl-2 protein expression in laryngeal squamous cell carcinoma and to clarify the relationship between them. All patients with laryngeal carcinoma were treated during the period 1991–1993. In the present study, p53 and bcl-2 expression in paraffin sections from 50 cases of laryngeal squamous cell carcinoma were analysed and correlated with routine clinico-pathological parameters. The expressions of p53 and bcl-2 were examined by immunohistochemical staining. Immunoreactivity for p53 was observed in 45 (90%) of carcinomas and bcl-2 immunoreactivity in 7 (14%). No significant correlation between the p53 or bcl-2 expression and patients’ T- or N-stage, histological grading, or overall survival was found. Received: 2 May 2001 / Accepted: 7 June 2001     

Association of p53 expression with second primary tumour development in hypopharyngeal carcinoma

Abnormalities of p53 tumour suppressor gene are detected in a diversity of malignancies and play an important role in their pathogenesis. Hypopharyngeal carcinoma is the most morbid among head and neck squamous cell carcinomas because of the high incidence of treatment failures and because a biological marker predictive of the treatment failures remains elusive. The expression of p53 protein in 46 hypopharyngeal squamous cell carcinomas was examined histochemically and p53 immunoreactivity was found in 19 of 46 cases (41.3 per cent). The rate of second primary tumour development was significantly higher in the p53-positive group than in the p53-negative group (p = 0.039), whereas that of tumour recurrence was not significantly different between the two. Moreover, there was no statistically significant difference in either overall or disease-free survival between the p53-positive and -negative groups. These results indicate that although p53 expression significantly correlates with second primary tumour development in patients with hypopharyngeal squamous cell carcinomas, it is not predictive of the clinical outcome.     

Bcl-2蛋白在头颈恶性肿瘤中的表达

为了探讨Ｂｃｌ－２蛋白在头颈恶性肿瘤发生、发展中的作用及与抑癌基因Ｐ５３的关系，采用免疫组化链霉力素生物素技术（ＬＳＡＢ法），对９０例头颈恶性肿瘤和２０例良必焦组织中的Ｂｃｌ－２，ｐ５３蛋白进行了检测。结果显示，４８．９％的头颈恶性肿瘤表达Ｂｃｌ－２蛋白，而２０例良性病变全部阴性。Ｂｃｌ－２在头颈鳞癌的表达率（５９．６％）明显高于腺癌（３４．８％）、未分化癌（３３．３％）和恶性淋巴瘤。在Ｐ５３阳性     

p21WAF1/CIP1和抑制增殖细胞核抗原的表达与下咽癌及喉癌的临床关系

目的：探讨p21^WAF1/CIP1和抑制增殖细胞核抗原(PCNA)的表达与下咽癌、喉癌的分级、转移及预后的关系,了解p21^WAF1/CIP1在下咽癌、喉癌发生、发展中的作用以及与PCNA的相关性。方法：对45例下咽癌、25例声门上型喉癌、23例声门型喉癌标本,用Envision HIS的方法检测p21^WAF1/CIP1、PCNA的表达情况,并分析p21^WAF1/CIP1表达与下咽癌、喉癌临床病理特征、转移、复发和预后的关系。结果：p21^WAF1/CIP1阳性表达率为51.6％(48／93),PCNA的阳性表达率为97.8％(91／93);p21^WAF1/CIP1表达与下咽癌、喉癌患者的年龄、性别、肿瘤分化、远处转移及复发无显著性相关,与肿瘤的原发部位的T分期有显著性相关;p2^WAF1/CIP1的表达还与肿瘤的预后有显著性相关;而PCNA与肿瘤的临床分期及淋巴转移有显著性相关,p21^WAF1/CIP1与PCNA的表达具有相关性。结论：p21^WAF1/CIP1、PCNA是预测肿瘤浸润范围及转移的良好指标,p21^WAF1/CIP1对预后的判定具有一定的指导意义。     

Telomerase catalytic subunit gene expression does not influence survival of patients with squamous cell carcinoma of the larynx and hypopharynx: a case-control study

AIMS: to determine correlations between relative quantities of telomerase catalytic subunit m-ribonucleic acid (hTERT mRNA) and conventional clinicopathological parameters (such as site, size and grade of tumour, the presence of regional lymph node metastases, and, in particular, survival) in patients with laryngeal and hypopharyngeal squamous cell carcinomas (SCCs). MATERIAL AND METHODS: The relative quantity of hTERT mRNA was analysed by a commercially available LightCycler Telo TAGGG hTERT Quantification Kit in 56 cases of SCC (40 laryngeal and 16 hypopharyngeal). The association with cancer-specific survival was evaluated by univariate and multivariate analysis. RESULTS: Location of the tumour in the hypopharynx was the only significant negative predictive factor for survival, as determined by univariate analysis (p = 0.028). Although a tendency towards a better overall survival was observed for female patients younger than 50 years, for lower tumour grades and sizes, and for the absence of regional lymph node metastases, the prognostic significance of these factors could not be confirmed. No differences existed in hTERT mRNA expression between laryngeal and hypopharyngeal SCCs. Furthermore, no correlation was found between the relative quantities of hTERT mRNA and the tumour size, regional lymph node metastases or survival of patients with laryngeal or hypopharyngeal SCCs. CONCLUSIONS: The results of the present study suggest that genetic abnormalities other than telomerase reactivation are responsible for progression of laryngeal and hypopharyngeal SCCs.     

Vascular endothelial growth factor expression correlates with p53 mutation and angiogenesis in squamous cell carcinoma of the head and neck

Vascular endothelial growth factor (VEGF) has potent angiogenic activity and has been identified in a wide variety of malignancies, including head and neck squamous cell carcinoma (HNSCC). The tumour-suppressor gene p53 has been thought to regulate VEGF. Cryostat sections of 33 head and neck squamous cell carcinomas (HNSCC) were immunostained for VEGF using a standard streptavidin-biotin complex procedure. To evaluate angiogenesis, microvascular density was counted by staining endothelial cells immunohistochemically using anti-vWF monoclonal antibody. The p53 gene status was analysed using a PCR-SSCP analysis and direct sequencing. VEGF positive staining was detected in 18 (55%) out of 33 tumours. VEGF immunoreactivity did not correlate with the main clinicopathological characteristics of the patients (localization, T-stage, N-status, histological grading). Statistical analysis gave a clear correlation between the tumour vascularity and the VEGF protein expression (p = 0.0036). VEGF negative tumours showed a lower mean number of microvessels per microscopic field (60.3 +/- 15.5) than VEGF positive tumours (79.6 +/- 22.9). P53 mutations were identified in 12 (36.4%) of 33 tumours. The association of p53 mutations and VEGF expression level was significant (0.027). The higher microvessel density in VEGF positive tumours supports the importance of VEGF for tumour angiogenesis in HNSCC. Our results support the hypothesis of a p53 regulation on the angiogenic process through a VEGF up-regulation.     

头颈部鳞癌颈淋巴结转移诸因素对预后的影响

对382例头颈部原发鳞癌病人颈清扫标本连续切片病理观察结果和临床资料进行回顾性分析，探讨淋巴结转移的各项临床和病理学因素与病人预后的关系。发现总体5年生存率为46.1％，口腔癌、口咽癌、下咽癌和喉癌的5年生存率分别为49．7％，39．7％，35．0％和60．3％。表明颈淋巴结的临床分期、触诊淋巴结大小、病理转移淋巴结情况、颈淋巴解剖分区受累数和最低受累平面与病人5年生存率有密切关系（均P＜0．01），而阳性淋巴的个数对病人预后无明显影响。提示针对影响预后的淋巴结因素应采取相应的治疗措施，以提高病人的5年生存率。     

Over-expression of tumour suppressor gene p53 in laryngeal squamous cell carcinomas and its prognostic significance

p53 is a nuclear phosphoprotein which acts as a tumour suppressor factor, regulating cell growth and division. Mutations in the p53 gene appear to be the most common genetic alterations in human cancer. The aim of this study was to investigate p53 expression in laryngeal squamous cell carcinomas and to assess its role as a marker of prognostic significance. Using immunohistochemical staining techniques, a series of laryngeal carcinomas (n= 87) were examined for expression of the mutant form of p53 phosphoprotein using the monoclonal antibody PAB 1801. p53 over-expression was noted in 50 biopsies of laryngeal carcinomas (57.5%) but not in any of the non-neoplastic laryngeal mucosa which were used as the control. There was no statistical correlation between p53 immunoreactivity and the clinicopathological parameters of the cancers including: site of tumour, TNM staging, differentiation grading and tumour recurrence. These findings indicate that p53 expression is strongly associated with carcinoma cells and not with normal cells in the larynx. However, p53 expression is probably unrelated to the biological aggressiveness of these tumours.     

Lymphatic vessel density, nodal metastases, and prognosis in patients with head and neck cancer

OBJECTIVE: To examine the relationship between intratumoral lymphatic vessel density and clinical and pathological variables in patients with head and neck squamous cell carcinoma. DESIGN: Archived paraffin-embedded biopsy specimens were sectioned and stained with hematoxylin-eosin and anti-LYVE-1 antibody, a highly specific marker for lymphatic endothelium. Tumor grade, infiltrating margin, inflammatory infiltrate, and percentage of tumor necrosis were noted and lymphatic vessel density measured using Chalkley point counting. SETTING: Tertiary care center at a university hospital. PATIENTS: A total of 168 previously untreated patients with advanced squamous cell carcinoma (73, larynx; 62, oropharynx; and 33, hypopharynx) treated with primary radiation (with or without planned neck dissection) and salvage surgery from 1992 to 1999. INTERVENTIONS: Measurement of intratumoral lymphatic vessel density in pretreatment tissue biopsy specimen. MAIN OUTCOME MEASURES: Disease-free and disease-specific survival, tumor occurrence, and nodal status. RESULTS: In patients with laryngeal carcinoma there was a significant relationship between the presence of intratumoral lymphatics and nodal metastases at presentation (P = .02) and poorly differentiated tumor grade (P = .02). Patients with high lymphatic vessel density also had a significantly worse disease-specific survival (P = .03). However, this difference was not significant with multivariate analysis. No significant relationship existed between the presence of intratumoral lymphatics and any of the clinical or pathological variables in oropharyngeal and hypopharyngeal carcinoma. CONCLUSIONS: In this patient sample, the development of intratumoral lymphatics in laryngeal carcinoma, but not in oropharyngeal or hypopharyngeal carcinoma, is associated with a spread of the tumor to regional lymph nodes. Detecting tumor lymphatic vessel proliferation is another step in the understanding of tumor biology, and the targeting of lymphatic growth may be of potential therapeutic benefit in selected patients with head and neck squamous cell carcinoma.     

Cell kinetics and apoptosis in laryngeal carcinoma patients

Cellular proliferation and apoptosis are both implicated in the process of carcinogenesis. The objective of this study was to access the prognostic significance of the expression of proliferating cell nuclear antigen (PCNA) and the apoptosis-related genes (bax, bcl-2, and p53) in laryngeal carcinoma patients. Thirty consecutive patients with stage I to IV squamous cell laryngeal carcinoma were treated in our department from 1992 to 1994. We immunohistochemically studied the expression of PCNA and bax, bcl-2, and p53 genes in their tumor specimens. Five healthy men were used as the control group. The staining results were correlated with clinicopathologic data. The PCNA protein expression was correlated with a significantly worse survival in those patients who were bax-negative (0% versus 42.86%, p = .0445). Similarly, the presence of PCNA led to an unfavorable clinical outcome in those patients who were bax-negative, bcl-2-negative, and p53-negative (0% versus 50%, p = .0278). Expression of bcl-2 protein was found to be an independent prognostic factor related to an unfavorable clinical outcome (p = .0262). The expression of bcl-2 protein appears to predict survival in laryngeal carcinoma patients. Furthermore, the combined study of proliferation markers and apoptosis-related genes helped us to identify a high-risk group of patients who may benefit from a more aggressive treatment protocol.     

An immunohistochemical analysis of p53 protein expression in pre-malignant and malignant tissues of the oral cavity

This study looks at p53 protein expression in dysplastic and malignant lesions of the oral cavity using an immunohistochemical staining technique. Archival biopsy specimens of oral dysplasia of squamous cell carcinoma from 64 patients were analysed immunohistochemically. Sections from 90 oral biopsy specimens were examined in all. Positive immunohistochemical detection of the p53 protein, demonstrated by brown nuclear staining, was detectable in over 80% of mild, moderate and severe dysplastic tissues as well as carcinoma-in-situ and squamous carcinoma specimens. We concluded that p53 protein expression occurs frequently in both malignant and dysplastic lesions of the oral cavity, suggesting that abnormally detectable p53 protein is present at the very early stages of development of oral squamous carcinoma. Oral cancer may provide a good model for the study of multistage tumorigenesis in head and neck cancer as the lesions are frequently detected at the pre-invasive stage and are accessible to biopsy     

survivin在喉癌和下咽癌中的表达及与p53、bcl-2蛋白表达的关系

目的:探讨survivin在喉癌和下咽癌中的表达情况及与p53、bcl-2间的相互关系.方法:采用免疫组织化学SP法榆测40例喉及下咽癌和10例声带息肉组织中survivin、p53及bcl-2的表达.结果:survivin、p53及bcl-2在喉癌及下咽癌中的阳性表达率分别为70.0%、57.5%和37.5%,而三者在声带息肉组织中均无表达(P＜0.05).survivin的表达随着病理分化程度的降低而增加,与临床分期、淋巴结转移成正相关(P＜0.05),与年龄无关;bcl-2的表达与临床分期成负相关(P＜0.05),与其他参数无关;p53的表达与各临床病理参数均无明显相关性.survivin在喉癌和下咽癌中的表达与p53的表达成正相关(P＜0.05),而与bcl-2的表达无相关性(P＞0.05).结论:survivin、p53可能参与喉癌及下咽癌的发生、发展并在其中起协同作用,但bcl-2则可能是通过不同的机制参与喉癌及下咽癌的发生.     

71例头颈部鳞状细胞癌的多原发癌临床资料分析

目的　了解头颈部鳞癌的多原发癌发病部位和治疗、生存状况。方法　对 71例头颈部鳞状细胞癌 (简称鳞癌 )的多原发癌临床资料做了回顾性分析。结果　发生在头颈部的重复癌 2 7例 ,发生在非头颈部的重复癌 4 2例 ,另有三重癌 2例。本组共有同时性重复癌 4例 ,其中 1例是同时性三重癌。余 6 7例均为异时性重复癌 ,其中 1例为异时性三重癌。 6 7例异时性重复癌中 ,先证癌与重复癌发生的间隔期为 8个月～ 12年不等。 70 %的先证口腔鳞癌发生头颈部重复癌 ;6 2 %的先证下咽癌和 79%的先证喉癌发生非头颈部位的重复癌。非头颈部位以食管和肺部发生的重复癌较多。在所有头颈部鳞癌发生重复癌的部位中 ,以食管重复癌为最多 ,占本组病例的 2 4 %。本组病例总体3年、5年生存率分别为 32 4 %和 2 2 5 % ;重复癌治疗组和未治组的 3年生存率有明显统计学差异 ,治疗组明显高于未治组。结论　头颈鳞癌的重复癌以食管癌最为多见。口腔癌容易发生头颈部重复癌 ,喉癌和下咽癌易发生非头颈部重复癌。细致随访和复查、早期明确诊断和积极有效的治疗 ,可以提高这类患者的生存率。     

Possible prognostic role of p53 expression in laryngeal squamous cell carcinoma of non-smokers,non-alcoholic patients

Conclusions. p53 expression seems to negatively influence survival in non-smoking non-alcoholic patients with squamous cell laryngeal carcinoma. p53 might be implicated in the oncogenic pathways leading to neoplastic transformation in this population of patients. Objectives. The aim of this study was to analyze the prognostic significance of p53 immunohistochemical expression in laryngeal squamous cell carcinoma arising in non-smoking non-alcoholic patients. Patients and methods. We retrospectively reviewed the data from 782 patients diagnosed with laryngeal squamous cell carcinoma in the last 15 years at Mostoles Hospital. From these patients we chose 21 who had never smoked or drunk alcohol and performed immunohistochemical staining for p53 protein in paraffin-embedded archival blocks. Results. We demonstrated immunohistochemical positivity for p53 in the samples from eight patients, most of them with a focal and weak pattern (<25% positivity in six cases). p53 expression did not show any significant prognostic influence on the risk of local or regional recurrence of the tumor, but significantly influenced the risk of death (log-rank test, p=0.03).     

喉乳头状瘤临床行为与细胞增殖活性关系的研究

利用ISH方法以及免疫病理手段对不同型的LP组织的标本进行PCNA、P53检测，探讨不同型HPV与LP的发生发展的相关机制，以寻求有效的检测方法帮助临床对LP的预后进行评估。标本选自1994年1月～1995年12月间我院收治的LP共36例。ISH法HPV6b／11阳性率为75％，明显高于HPV16和／或HPV18的表达。10例喉鳞癌各有1例HPV16、18阳性，无HPV6b／11阳性。ABC法行P53检测，36例LP标本中仅1例恶变组织阳性表达Ⅱ级（2．8％）；10例喉鳞癌中9例阳性表达（90．0％），其中Ⅱ级以上阳性表达6例（60．0％）。PCNA阳性表达27／36例（75％）；其中JOP组与AOP组阳性率有显著差异（P＜0．05）。本研究表明LP与HPV感染有极为密切的关系，认为HPV分型的检测在判断LP转归中有意义。PCNA阳性表达程度在预测LP肿瘤的活跃程度方面是一个很有意义的指标。P53蛋白表达在喉鳞癌与LP中有显著差异。     

p53 and cyclin D1 as prognostic factors in squamous cell carcinoma of the larynx

OBJECTIVES/HYPOTHESIS: 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. STUDY DESIGN: Retrospective multicentric study. METHODS: 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. RESULTS: 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. CONCLUSION: 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     

Características diferenciales de los pacientes con carcinomas escamosos de cabeza y cuello en función del género

ObjectiveTo analyse the trend in the percentage of patients with squamous cell carcinoma of the head and neck of female gender over the last decades, and differences in survival according to gender.Material and methodsRetrospective study of 5,828 squamous cell carcinoma of the head and neck diagnosed during the period 1985-2019. In the survival analysis we only included only patients with more than two years of follow-up.ResultsThe proportion of tumours in female patients increased significantly over the study period. There was greater increase of tumours located in the oral cavity, oropharynx and larynx in the percentage of female patients. When considering the total number of patients, there were no significant differences in disease-specific survival according to gender. We observed a variable behaviour of disease-specific survival according to gender depending on the primary location of the tumour. For tumours of the larynx and oropharynx the female patients had a disease-specific survival significantly higher than that of men, while for tumours of the oral cavity, the female patients had a significant reduction of disease-specific survival.ConclusionsIn our setting, the percentage of female patients with squamous cell carcinoma of the head and neck has been increasing steadily over recent decades. There were differences in disease-specific survival according to gender depending on the location of the primary tumour.     

p53蛋白异常表达与喉癌,下咽癌颈淋巴结转移的相关研究

应用免疫组化方法（ＬＳＡＢ法）对５７例喉癌、下咽癌标本ｐ５３蛋白染色，进行颈淋巴结转移的相关性研究，结果发现：１）ｐ５３蛋白表达与肿瘤的临床分期有良好的对应关系，即随着临床分期的升高，ｐ５３蛋白表达率升高（Ｐ＜０．００１）；２）随肿瘤分化程度的降低，ｐ５３蛋白表达率升高（Ｐ＜０．００１）；３）ｐ５３蛋白在淋巴结转移组的表达明显高于无淋巴结转移组（Ｐ＜０．００１）。提示ｐ５３蛋白基因的突变对喉癌、下咽癌的颈淋巴转移有促进作用，因此，检测ｐ５３蛋白的表达可作为临床预测喉癌、下咽癌颈淋巴结转移趋势的重要参考指标之一。