Expression of E-cadherin, and CD44s and CD44v6 and its association with prognosis in head and neck cancer

OBJECTIVES: In the current study, the expression of E-cadherin, CD44s, and CD44v6 has been noted as markers for tumor metastasis and prognosis in several tumors, so we examined whether or not E-cadherin, CD44s, and CD44v6 are useful markers for evaluating the prognosis of mesopharyngeal cancer patients. METHODS: The expression of E-cadherin, CD44s, and CD44v6, was evaluated immunohistochemically using monoclonal antibodies against epitopes of standard and variant proteins, in paraffin-embedded mesopharyngeal cancer tissues from 57 patients who had received curative therapy. RESULTS: Tumor tissues from 47 (82.5%) patients showed positive immunoreactivity with monoclonal antibody against E-cadherin, 43 (75.4%) patients showed positive expression with CD44, and 45 (78.9%) patients showed positive expression with CD44v6. The expression of CD44v6 was slightly correlated with tumor volume, and lymph node metastasis, and stage classification (P > 0.05). However, there was no significant correlation between the expression of E-cadherin, CD44s and CD44v6 and clinicopathological characteristics. Concerning the prognosis, the survival period of patients with CD44s positive tumors was shorter than that of patients with CD44s negative tumors (18.2% versus 52.1%, 5-year survival, P > 0.05). The survival period of patients with CD44v6 positive tumors was also shorter than that of patients with CD44v6 negative tumors (12.8% versus 55.6%, 5-year survival, P > 0.05). CONCLUSION: These results suggest that CD44v6 may be related to tumor invasion and metastasis, and both CD44s and CD44v6 may be useful markers for poor prognosis in head and neck cancer.     

Two sites of head and neck squamous cell carcinoma: utility of loss of heterozygosity

OBJECTIVES: The prevalence of distant metastasis and second primary tumors is increasing with improved locoregional control in patients with head and neck squamous cell carcinoma. Traditionally, clinicopathologic evidence has been the gold standard used to distinguish distant metastasis from second primary tumors. We report a case in which loss of heterozygosity testing was used to clarify the clonal relationship between the 2 sites of head and neck squamous cell carcinoma. METHODS: A patient with squamous cell carcinoma in the larynx and mandible underwent loss of heterozygosity testing. RESULTS: The loss of heterozygosity testing confirmed that the mandibular cancer was a metastatic presentation of the laryngeal squamous cell carcinoma. CONCLUSIONS: We conclude that loss of heterozygosity testing can be useful in differentiating distant metastasis from second primary cancers in patients with 2 sites of head and neck squamous cell carcinoma, consequently providing important prognostic and staging information.     

Molecular analysis of surgical margins in head and neck squamous cell carcinoma patients

OBJECTIVES/HYPOTHESIS: Molecular analysis of surgical margins is playing an increasingly important role in establishing surgical margins. Most markers lack the sensitivity and ease of applicability for effective clinical use. To date, the proto-oncogene eIF4E (4E) is elevated in 100% of head and neck squamous cell carcinoma tumors and is of prognostic value in predicting recurrence. In a retrospective study, 4E overexpression in the margins appeared to be a more sensitive predictor of recurrence when compared with p53. The goal was to confirm this finding in a prospective study and also to compare the expression of matrix metalloproteinase-9 (MMP-9) to 4E expression in tumors and margins. Other objectives were to determine which of these markers have prognostic significance in predicting recurrence and elucidate whether there is any additional benefit to analysis of surgical margins with a combination of the three molecular markers. STUDY DESIGN: A prospective study was performed on all patients who consecutively underwent primary surgical resection between 1998 and 1999 for head and neck squamous cell carcinoma. Patient and tumor characteristics were reviewed, and time to recurrence was noted. METHODS: Paraffin-embedded sections of tumors and all histologically tumor-free margins were analyzed for the presence or absence of 4E, p53, and MMP-9 with immunohistochemical analysis. Patients were followed according to the institution's head and neck cancer protocol, and time to recurrence was noted. RESULTS: Ninety-eight percent of tumors overexpressed 4E, 65% overexpressed p53, and 92% overexpressed MMP-9. Of the 52 patients with tumor-free margins, 52%, 46%, and 54% had positive margins for 4E, p53, and MMP-9, respectively. Although no significant correlation between 4E and p53 expression was seen in the margins (P =.16), a significant correlation between 4E and MMP-9 expression was noted (P =.0002). However, when expression of 4E and p53 in the margins of only the patients who overexpressed p53 in the tumors was compared (n = 34), there was a significant correlation (P =.04). There was also a significant difference in the disease-free interval between patients with 4E-positive and 4E-negative margins (P =.003). This difference in time to recurrence was not significant for the p53-positive versus the p53-negative group (P =.18) but approached significance when MMP-9 was used as a marker (P =.07). Although the univariate analysis showed that stage, nodal disease, grade, and 4E expression in the margins were significantly associated with disease-free interval, in the Cox multiple regression analysis, only 4E expression in the margin was significantly associated with disease-free interval (P =.01). CONCLUSIONS: The era for molecular analysis of surgical margins is here. Although a significant correlation was seen between 4E and MMP-9, overexpression of 4E appears to be a significant predictor of recurrence when compared with the well-studied tumor suppressor gene p53 and a relatively novel marker, MMP-9.     

Molecular pathways of lymphangiogenesis and lymph node metastasis in head and neck cancer

Metastasis to regional lymph nodes constitutes the main route toward progression and dissemination of head and neck carcinoma; at the same time it is the most significant adverse prognostic indicator for this disease. In recent years, significant focus has been given on the molecular mechanisms behind lymph node metastasis of head and neck cancer. The aim of this study is to assess the role of growth factor expression and function in association with lymph node metastasis and overall prognosis of head and neck cancer. Current literature, searching for experimental data regarding the molecular pathways of lymph node dissemination of head and neck cancer, is reviewed giving special emphasis on the expression and prognostic significance of specific growth factors. Members of the vascular endothelial growth factor (VEGF), mostly VEGF-C and VEGF-D, with their action through the receptors VEGFR-3 and VEGFR-2, constitute the most extensively studied growth factors associated with lymphangiogenesis so far. High expression of these as well as other molecules, including angiopoietins, insulin-like growth factor, and fibroblast growth factor, has been associated with lymph node metastasis and poor prognosis in head and neck squamous cell carcinoma. Numerous growth factors seem to play an important role regarding the lymph node metastatic potential of head and neck cancer. Further research is necessary in order to further clarify the molecular pathways and introduce novel therapeutic options.     

Focal adhesion kinase and E-cadherin as markers for nodal metastasis in laryngeal cancer

OBJECTIVE: To explore the value of E-cadherin and focal adhesion kinase (FAK) expression in the prediction of cervical lymph node metastases in squamous cell carcinoma of the supraglottic larynx. DESIGN: Immunohistochemical analysis of retrospectively selected cases. Patients The study population was composed of 95 previously untreated men with squamous cell carcinoma of the supraglottic larynx. Intervention All the patients underwent surgical resection of the tumor and bilateral neck dissection. MAIN OUTCOME MEASURES: E-cadherin and FAK expression in relation to nodal metastases. RESULTS: Decreased E-cadherin expression was correlated with the presence of nodal metastases (P = .006). The combination of E-cadherin and FAK expression resulted in a superior accuracy in assessing nodal metastasis (P = .001). Histological grade was also associated with nodal metastases (P = .005). Multivariate analysis confirmed that these parameters were independent predictors of nodal metastases. In addition, the cases with decreased E-cadherin and increased FAK expression presented a significantly reduced disease-specific survival (P = .005). CONCLUSION: The combination of the expression of E-cadherin and FAK could increase our ability to identify patients with clinically negative lymph nodes who are at considerable risk for occult metastases.     

Expression of the E-cadherin-catenin cell-adhesion complex and CD44 variant 6 in primary squamous cell carcinomas of the head and neck and their nodal metastases

Aims: A reduction in cell–cell adhesion followed by cell invasion are essential steps in the progression from localized malignancy to metastatic disease. In this study, changes in the expression of the components of the E-cadherin-catenin cell-adhesion complex and CD44v6 have been investigated. Methods: The above molecules were studied using immunohistochemical techniques in fresh sections of primary tumours and nodal metastases from 36 patients with squamous cell carcinoma of the head and neck. For 14 patients the corresponding primary and nodal metastases samples were available. Results: All 51 samples showed reduced E-cadherin expression when compared with either the adjacent normal squamous epithelium or with normal controls. Expression of β-catenin generally parallelled that of E-cadherin and expression of α-catenin was generally weak. For CD44v6, eight primary tumours exhibited membranous labelling comparable to the normal controls while the remainder showed reduced membranous and increased cytoplasmic labelling. The nodal metastases showed fewer cells with membranous labelling with all the antibodies, than the primary carcinomas. Marked intratumoural heterogeneity for protein expression was evident for all antibodies, and the abnormal expression of the catenins is a novel finding. E-cadherin was expressed more intensely in cells with greater squamous differentiation and there was a correlation between CD44v6 expression and T-stage. There was no correlation between decreased expression of any of the adhesion molecules and local recurrence, metastasis, or survival. Conclusion: The loss of expression of E-cadherin and decrease in CD44v6 may be permissive factors for metastasis in squamous carcinoma of the head and neck, but they do not appear to be the only determinants of this process.     

Prognostic significance of p53 and FHIT in advanced oropharyngeal carcinoma

OBJECTIVE: To determine the prognostic significance of p53 and fragile histidine triad (FHIT) expression in advanced oropharyngeal squamous cell carcinoma. STUDY DESIGN: A retrospective collection of clinical data was correlated with the protein expression. METHOD: The expression of p53 and FHIT in specimens from patients with previously untreated advanced squamous cell carcinoma of the oropharynx was determined by immunohistochemistry. The expression of p53 and FHIT was statistically correlated with survival outcome. The primary endpoints were overall survival and disease-free survival. RESULTS: Thirty-four patients were analyzed in this study. Overexpression of p53 was observed in 41.2% (14/34) of tumors and was associated with a trend toward an improved overall survival using univariate (P =.1088, risk ratio [RR] = 0.503) and multivariate (P =.1533, RR = 0.470) analyses. Marked reduction or complete absence of FHIT expression was observed in 57.6% (19/33) of tumors. Patients with tumors showing no reduction in FHIT expression had a lower overall survival using univariate (P =.04, RR = 2.27) and multivariate (P =.013, RR = 4.41) analyses. CONCLUSION: Overexpression of p53 predicted a trend toward an improved prognosis, whereas no reduction in FHIT expression predicted a significantly poorer outcome in patients with advanced oropharyngeal cancer.     

Prognostic factors in head and neck mucoepidermoid carcinoma

OBJECTIVE: To analyze clinical, histological, and immunohistochemical prognostic factors in a large series of patients with mucoepidermoid carcinoma (MEC) treated in a single institution, using univariate and multivariate survival analyses. DESIGN: Inception cohort. SETTING: Referral center. PATIENTS: All patients diagnosed with head and neck MEC from a single cancer referral center from January 19, 1957, to July 12, 1997. MAIN OUTCOME MEASURES: Rates of local recurrence, regional and distant metastasis, and overall actuarial survival. RESULTS: Men represented 53.8% of the cohort, and the parotid gland and palate were affected by MEC in 35.2% and 23.7%, respectively. TNM stage I or II lesions comprised 50.3% of the tumors, and low-grade tumors comprised 45.2%, and the 5-year overall survival was 70.2%. Univariate survival analysis revealed that age older than 40 years (P<.001), male sex (P=.005), fixed tumors (P=.002), invasion of adjacent structures (P=.004), T stage (P<.001), N stage (P<.001), clinical stage (P<.001), histological grade (P<.001), and expression of proliferating cell nuclear antigen (P<.001), Ki-67 (P<.001), and p53 (P<.001) correlated with a poor prognosis. Expression of carcinoembryonic antigen (P=.01) and bcl-2 (P<.001) correlated with a better prognosis. CONCLUSION: Age older than 40 years, fixed tumors, T and N stage, and histological grade are independent significant prognostic factors in patients with MEC.     

Radiotherapy alone for clinical T4 skin carcinoma of the head and neck with surgery reserved for salvage

PURPOSE: To evaluate the outcomes of definitive radiotherapy in the treatment of clinical stage T4 cutaneous carcinomas of the head and neck. PATIENTS AND METHODS: Between October 1964 and September 1997, 85 patients with 88 biopsy-proven clinical AJCC stage T4 carcinomas of the skin of the head and neck received radiotherapy with curative intent. A total of 43 lesions were previously untreated, and 45 were recurrent after other treatment modalities. Histologic types of carcinoma included squamous cell (37 lesions), basal cell (41 lesions), and metatypical basal (basosquamous) cell (10 lesions). Minimum follow-up was 2 years. The product-limit method was used to determine the rates of disease control, severe late complications, and survival. Multivariate analyses included histology, previous treatment, involvement of bone or nerve, number of structures invaded, node stage, external beam dose, and overall treatment time. RESULTS: At 5 years, the rates of local control after radiotherapy and ultimate local control after salvage surgery were 53% and 90%, respectively. Local control rates were better for patients having previously untreated lesions (P =.05). Regional and ultimate regional control rates were 93% and 100%, respectively, and were better for previously untreated lesions (P <.01), basal cell histology or its metatypical variant (P =.04), and absence of bone invasion (P =.08). At 5 years, the risk of severe late complications was 17%, the risk of distant metastasis was 5%, and the overall absolute and cause-specific survival probabilities were 56% and 76%, respectively. CONCLUSION: Radiotherapy alone results in a relatively high probability of cure for selected patients with T4 skin cancers.     

Motility-related proteins as markers for head and neck squamous cell cancer

HYPOTHESIS: Increased cell motility is a hallmark of cancer cells. Proteins involved in cell motility may be used as molecular markers to characterize the malignant potential of tumors. METHODS: Molecular biology and immunohistochemistry techniques were used to investigate the expression of a selected panel of motility-related proteins (Rho A, Rac 2, Cdc42, PI3K, 2E4, and Arp2) in normal, premalignant, and squamous cell cancer cell lines of human head and neck origin. To assess the clinical potential of these proteins as molecular markers for cancer, immunohistochemistry was performed on paraffin-fixed head and neck cancer specimens (n = 15). RESULTS: All six motility-associated proteins were overexpressed in the premalignant and squamous cell cancer cell lines relative to normal keratinocytes. Immunohistochemistry with Rho A and Rac 2 showed increased staining in areas of cancer but not in normal tissue. CONCLUSION: Proteins involved in cell motility can be used as markers for head and neck squamous cell carcinoma. The head and neck cell lines used in this study may be used as a model to further investigate cell motility. Molecular markers of motility could have a significant impact on the diagnosis and staging of cancers originating from differentiated non-motile cells.     

Expression of E-cadherin, CD44s, and CD44v6 in laryngeal and pharyngeal carcinomas

OBJECTIVE: Tumors arising from different sites of the head and neck area have very different clinical behavior. Loss or reduction of expression of adhesion molecules has been assumed to play a critical role in the development of head and neck carcinomas. The aim of this study is to determine if there are differences in the expression of adhesion molecules E-cadherin, CD44s, and CD44v6 in pharyngeal and laryngeal squamous-cell carcinomas. MATERIALS AND METHODS: E-cadherin, CD44s, and CD44v6 expression was determined by immunohistochemistry in paraffin-embedded tissue specimens from 72 patients with squamous-cell carcinoma, 37 of the pharynx and 35 of the larynx. RESULTS: Expression of CD44s was significantly lower in pharyngeal than in laryngeal tumors (P =.01). No differences in the expression of E-cadherin and CD44v6 were observed between these sites. CONCLUSIONS: These data suggest that there are some differences at molecular level between the different subsites of head and neck cancer.     

头颈鳞癌血管生成与颈淋巴结转移相关性研究

目的 :探讨头颈鳞癌血管生成与其颈淋巴结转移的关系以及血管内皮生长因子(VEGF)在头颈鳞癌血管生成中的作用。方法 :应用免疫组化SABC法检测 5 8例头颈鳞癌组织中微血管密度 (IMVD)及VEGF的表达。结果 :5 8例头颈鳞癌组织中IMVD为 2 3.93± 8.77,肿瘤分化程度 ,高与中、高与低间 ,IMVD差异有显著性意义 (均P <0 .0 5 ) ;中与低间 ,差异无显著性意义 (P >0 .0 5 )。颈淋巴结转移组IMVD(2 7.92± 9.11)明显高于非转移组 (2 0 .6 9± 7.0 8) ,其差异有显著性意义 (P <0 .0 1)。癌组织中VEGF表达与瘤内IMVD呈正相关 (rs=0 .4 87,P <0 .0 1)。结论 :瘤内IMVD可作为预测头颈鳞癌颈淋巴结转移的一个重要指标 ;VEGF可促进头颈鳞癌血管生成。     

Delayed regional metastases, distant metastases, and second primary malignancies in squamous cell carcinomas of the larynx and hypopharynx

OBJECTIVE: To determine the impact of delayed regional metastases, distant metastases, and second primary tumors on the therapeutic outcomes in squamous cell carcinomas of the larynx and hypopharynx. STUDY DESIGN: Chart review and statistical analysis. METHODS: A retrospective tumor registry analysis was made of patients with squamous cell carcinomas of the larynx and hypopharynx who were treated with curative intent in the Department of Otolaryngology-Head and Neck Surgery and the Radiation Oncology Center of the Washington University School of Medicine (St. Louis, MO) between January 1971 and December 1991 and developed delayed regional metastases (2 y after treatment), distant metastases, and second primary malignancies. RESULTS: In 2550 patients, the mean age (59.8 y), sex (8.5 male patients and 1 female patient), and tumor differentiation did not affect the incidence of delayed distant, regional, or second primary malignancies. The overall incidence of delayed regional metastases was 12.4% (317/2550 patients); distant metastases, 8.5% (217/2550); and second primary tumors, 8.9% (228/2550), with a 5-year disease-specific survival of 41%, 6.4%, and 35%, respectively. Second primary malignancies were not statistically related to the origin of the primary tumor, tumor staging, or delayed regional and distant metastases (P =.98). Delayed regional metastases and distant metastases were related to advanced primary disease (T4 stage), lymph node metastases (node positive [N+]), tumor location (hypopharynx), and locoregional tumor recurrence (P < or =.028). Advanced regional metastases at initial diagnosis (N2 and N3 disease) increased the incidence of delayed and distant metastases threefold (P =.017). These two metastatic parameters were significantly greater in hypopharyngeal tumors than in laryngeal tumors (P =.037). The incidences of delayed regional metastases by anatomical location of the primary tumor were as follows: glottic, 4.4%; supraglottic, 16%; subglottic, 11.5%; aryepiglottic fold, 21.9%; pyriform sinus, 31.1%; and posterior hypopharyngeal wall, 18.5%. The incidences of distant metastases were as follows: glottic, 4%; supraglottic, 3.7%; subglottic, 14%; aryepiglottic fold, 16%; pyriform fossa, 17.2%; and posterior hypopharyngeal wall, 17.6%. Seventeen hypopharyngeal tumors (2%) presented with M1 disease. Delayed regional metastases to the ipsilateral treated neck had a significantly worse survival prognosis than delayed metastases to the contralateral nontreated neck (P =.001). CONCLUSIONS: Conclusions are as follows: 1) The incidence of second primary tumors is independent from the primary tumor staging and distant and delayed regional metastases. The highest incidence occurred in patient groups with the highest disease-free survival rates (P =.0378). 2) Highest incidence of delayed and distant metastases occurred in hypopharyngeal tumors and was three times greater than in laryngeal cancers (P =.028). 3) Salvage therapeutic rates were poor for delayed metastases to the ipsilateral treated nodes and distant metastases as compared with contralateral neck metastases and second primary tumors (P =.001). 4) Delayed and distant lymph node metastases were significantly higher in advanced primary disease (T4 stage), locoregional recurrences, and regional disease (N2 and N3) (P =.028) in both the larynx and hypopharynx. 5) The higher incidence of delayed and distant metastatic disease was related to more advanced initial tumor presentation in hypopharyngeal cancer as compared with laryngeal cancer (P =.039). 6) Incidence of distant metastases was greatest between 1.5 and 6 years after initial treatment with a mean incidence being less than or equal to 3.2 years.     

Expression of the cell-cell adhesion molecule E-cadherin in squamous cell carcinoma of the head and neck

The expression of the cell-cell adhesion molecule E-cadherin has previously been shown to be reduced in poorly differentiated squamous cell carcinoma of the head and neck and absent in nodal metatases. Twenty-eight patients with previously untreated squamous cell carcinoma of the head and neck, 22 of whom had nodal metastses at presentation, were investigated for E-chdherin expression using the monoclonal antibody 6F9, specific for human E-cadherin. Reduced expression was seen in the poorly differentiated primary tumours, compared with well differented tumours, but this trend was not statistically significant. E-cadherin expression was present at reduced level in nodal metastases. It was also noted that, where both the primary tumour and corresponding nodal metastasis were investigated, E-cadherin expression was identical for both samples. The degree of E-cadherin expression did not correlate with survival. These data confirm a reduction in E-cadherin expression in poorly differentiated tumours. There was no correlation between E-cadherin expression and any of the host, tumour and treatment factors associated with malignacies of the head and neck region.     

乙酰肝素酶在头颈部鳞状细胞癌的表达及其临床意义

目的探讨乙酰肝素酶（heparanase，HPSE）在头颈部鳞癌的表达与临床病理特征及预后的关系。方法对62例头颈肿瘤手术切除标本进行检测，采用免疫组化技术检测HPSE在原发肿瘤组织及其淋巴转移灶中的表达，分析HPSE的表达同患者的年龄、临床分期、病理分级、有无淋巴转移及与预后的关系。结果HPSE在癌旁黏膜组织中不表达或少表达，在大多数头颈肿瘤组织中呈阳性表达，阳性表达率为69．3％（43／62），主要为胞质表达。HPSE的表达在患者的年龄、肿瘤的病理分级方面比较，差异无统计学意义（Х^2=0．05，Х^2=3．84，P值均〉0．05）；而在有无淋巴转移以及肿瘤的临床分期方面差异有统计学意义（Х^2=3．98，Х^2=8．06，P值均〈0．05）；HPSE在原发灶及其淋巴转移灶中的表达之间呈明显正相关（r=0．9162，P=0．001）；Kaplan—Meier法计算，HPSE阳性组的3年累积生存率为25．9％，HPSE阴性组的3年累积生存率为72．7％，二者差异有统计学意义（Х^2=11．607，P=0．001）。HPSE的表达和肿瘤的TNM临床分期分别为影响预后的独立因素，二者分别与患者的预后显著相关（Х^2值分别为16．86和19．73，P值均〈0．05）。结论HPSE在头颈鳞癌中表达显著增高。HPSE的表达与有无淋巴转移、肿瘤的临床分期密切相关。HPSE的表达、肿瘤的临床分期是影响患者预后的独立因素。     

Distributions of cervical lymph node metastases in oropharyngeal carcinoma: therapeutic implications for the N0 neck

OBJECTIVES: This study sought to investigate the patterns and distributions of lymph node metastases in oropharyngeal squamous cell carcinoma (SCC) and improve the rationale for elective treatment of N0 neck. MATERIALS AND METHODS: One hundred four patients with oropharyngeal SCC who underwent neck dissection between 1992 and 2003 were analyzed retrospectively. All patients had curative surgery as their initial treatment for the primary tumor and neck. A total of 161 neck dissections on both sides of the neck were performed. Therapeutic dissections were done in 71 and 5 necks and elective neck dissection was done on 33 and 52 necks on the ipsilateral and contralateral sides, respectively. Surgical treatment was followed by postoperative radiotherapy for 78 patients. The follow-up period ranged from 1 to 96 months (mean, 30 months). RESULTS: Of the 161 neck dissection specimens evaluated, 90 (56%) necks were found to have lymph node metastases found by pathologic examination. These consisted of 76 (73% of 104 necks) of the ipsilateral side and 14 (25% of 57 necks) of the contralateral side dissections. The occult metastatic rate was 24% (8 of 33) of ipsilateral neck samples and 21% (11 of 52) of contralateral neck samples. Of the 68 patients who had a therapeutic dissection on the ipsilateral side and had lymphatic metastasis, the incidence rate of level IV and level I metastasis was 37% (25 of 68) and 10% (7 of 68), respectively. Isolated metastasis to level IV occurred on the ipsilateral side in three patients. There were no cases of isolated ipsilateral level I pathologic involvement in an N-positive neck or occult metastasis to this group. The incidence rate of level IV metastasis in patients with ipsilateral nodal metastasis was significantly higher in base of tongue cancer (86% [6 of 7]) compared with tonsillar cancer (34% [20 of 59]) (P=.013). Patients with level IV metastasis had significantly worse 5-year disease-free survival rates than patients with metastasis to other neck levels (54% versus 71%; P=.04). CONCLUSION: These results suggest that elective N0 neck treatment in patients with oropharyngeal SCC, especially base of tongue cancer, should include neck levels II, III, and IV instead of levels I, II, and III.     

DNA ploidy of primary and metastatic squamous cell head and neck cancers.

Regional metastases are a major determinant in the treatment outcome of patients with squamous cell carcinoma of the head and neck. Metastases do not respond as well to cytotoxic therapy as do primary tumors. DNA diploid tumors or tumor components also respond poorly to intermittent cytotoxic therapy. In our series of 497 patients with squamous cell carcinoma of the head and neck, the percentage of pure DNA diploid tumors and the mean DNA indexes in 497 primary tumors and 82 regional metastases were 34% and 1.54 and 50% and 1.34, respectively. Paired comparisons were performed in 61 patients and revealed a statistically significant increase in the frequency of DNA diploid tumors (27.4% to 41.2%) in associated lymph node metastases. The clinical observation that patients with squamous cell carcinoma of the head and neck and regional lymph node metastases have a poorer prognosis and a poorer response to cytotoxic therapy may in part be explained by the increased incidence of DNA diploid tumors in their regional lymph nodes, and the poorer response of such tumors to cytotoxic therapy.     

Cyclin D1 amplification and p16(MTS1/CDK4I) deletion correlate with poor prognosis in head and neck tumors

OBJECTIVES/HYPOTHESIS: Cyclin D1, a cell cycle regulator localized to chromosome 11q13, is amplified in several human tumors including head and neck squamous cell carcinoma (HNSCC). Amplification and/or overexpression of cyclin D1 have been correlated to a poor prognosis. Deletion of the p16 gene, localized to 9p21, has also been observed in a significant proportion of HNSCC. The p16 gene regulates cyclin D1-CDK4 activity and prevents retinoblastoma tumor suppressor gene phosphorylation, thereby downregulating cellular proliferation. Detection of cyclin D1 amplification and p16 deletion using a simple and sensitive method will be valuable for the development of effective treatment modalities for head and neck cancer. STUDY DESIGN: We have used fluorescence in situ hybridization (FISH) to study cyclin D1 amplification and p16 gene deletion in head and neck tumors. Both single- and dual-color FISH were performed. METHODS: Paraffin-embedded tissues from 103 patients with HNSCC were analyzed using genomic DNA probes for cyclin D1 and p16. Dual-color FISH was performed with chromosome 11 or 9 centromeric probes as a control. Twenty-eight of these samples were analyzed for p16 expression by immunohistochemistry. RESULTS: Cyclin D1 amplification was observed in 30% (31/103) of patients, and p16 deletion in 52% (54/103). Lack of p16 expression was observed in 64% (18/28) of patients. There was a good correlation between the deletion of p16 sequences and the loss of p16 expression (P = .008). Amplification of cyclin D1 had a statistically significant association with recurrence, distant metastasis, and survival at 36 months. There was a significant association between p16 deletion and the development of distant metastases. Cyclin D1 amplification and p16 deletion together correlated with recurrence, distant metastasis, and survival. CONCLUSIONS: We demonstrate that FISH is a simple and sensitive method for detecting cyclin D1 amplification and p16 deletion in head and neck cancer. Our results suggest that these two genetic aberrations together portend a poorer outcome than either of the abnormalities alone in head and neck cancer.     

Lymph node metastases in the lower neck

Current knowledge suggests that lymph node metastases in the lower neck (supraclavicular fossa and posterior triangle) are associated with a poor survival. Very little systematic work has been published on this subject. This was a retrospective study carried out on a database where all patients were entered in a prospective manner over a 35-year period using a standard pro-forma. Data on 168 patients presenting with a lower neck node metastasis were retrieved. The main outcome measures were: association between variables and tumour-specific survival. Data were displayed in contingency tables and analysed by chi-square and categorical modelling. Recurrence and survival were plotted in a cause-specific manner using the Kaplan Meir method. Differences in curves were analysed using the log rank test. Multivariate analysis was carried out using Cox's proportional hazard model. The only association was between site and node level and histology. Head and neck tumours were associated with squamous histology (P = 0.0004) and supraclavicular nodes (P = 0.0047). Survival time was not significantly different when lower-neck lymph node metastasis from the head and neck was compared to non-head and neck metastasis: 5-year survival 30% and 10% respectively (P = 0.1363). Survival with posterior triangle metastases was significantly better than supraclavicular metastases (P = 0. 0059), confirmed on multivariate analysis. Laterality of metastasis had no effect on survival (P < 0.0001). There was no significant difference in survival between squamous and non-squamous metastases on Cox regression (P = not significant). There were 85 head and neck primaries including lymphomas, 53 infraclavicular primaries and 30 unknown primaries. There were 73 squamous cell carcinomas, 27 adenocarcinomas, 34 lymphomas, 28 undifferentiated tumours and six other tumours. Nearly half the primary tumours were below the clavicle. Survival was unaffected by laterality, primary site or histology, but was better for posterior triangle nodes.     

转化生长因子-1和上皮钙黏附素在声门上型喉癌中的表达与转移和预后的关系

目的：探讨转化生长因子-1（TGF-β1）和上皮钙黏附素（E-cd）在声门上型喉鳞状细胞癌（SGLSCC）中的表达与颈部淋巴结转移和预后的关系。方法：采用免疫组织化学SABC法对60例SGLSCC蜡块标本和10例正常喉黏膜标本中TGF-β1、E-cd的表达情况进行检测,结合临床及随访资料进行分析。结果：①60例SGLSCC中TGF-β1和E-cd的阳性表达率分别为81．7％（49／60）和70．0％（42／60）;10例正常喉黏膜中TGF-β1和E-cd的阳性表达率分别为20．0％（2／10）和90．0％（9／10）;②TGF-β1和E-cd在淋巴结、病理分级及临床分期3项在各组间的表达分别呈正、负相关关系;③两指标的表达与淋巴结转移有显著的关系,而与患者的年龄、性别及肿瘤大小无关;④两指标间无一致性但呈负相关关系;⑤E-cd的表达与生存率曲线有统计学意义。结论：SGLSCC中TGF-β1和E-cd的表达与淋巴结转移有关,两指标间呈负相关关系,两指标可作为预测SGLSCC淋巴结转移的标志物。而E-cd还可作为判断预后的标志物。