头颈部鳞癌及癌旁组织端粒酶活性检测

目的：研究原发头颈部鳞癌及相关癌旁组织中端粒酶活性表达，探讨春作为头颈部鳞癌分子生物学标志物的可能性。方法：采用ＴＲＡＰ－ＰＣＲ－ＥＬＩＳＡ，对３２例原发头颈部鳞癌及１５例癌旁组织进行端粒酶活性检测。结果：３２例原发头颈部鳞癌中，２７例端粒酶活化，阳性率为８４．４％；１５例癌旁组织中５例端粒酶活化，阳性率为３３．３％。有淋巴结累及者端粒酶阳性率（８６．７％）高于无淋巴结累及者（８２．４％），低分化     

癌旁非典型增生组织端粒酶活性的间接定量检测

目的对头颈部癌旁非典型增生组织中端粒酶活性进行间接定量测定;探讨该定量分析对预测癌旁非典型增生组织恶性变的价值.方法用定量方法检测取自10位头颈肿瘤病人的组织标本30份,其中原发癌组织、癌旁非典型增生组织及正常组织标本各10份.端粒酶活性检测采用端粒重复序列液体闪烁计数法并由此推断端粒酶活性.结果癌旁非典型增生组织中端粒酶活性(645±262) rcpm明显低于相应的癌组织(1618±329) rcpm,有统计学意义(P<0.01);高于相应的正常组织(506±209) rcpm,但差异无统计学意义(P>0.05).结论①端粒酶活性的液闪间接定量分析,对癌旁组织恶性变的预测可能有一定的参考价值,可用于肿瘤切缘的研究;②癌旁组织端粒酶活性升高进一步提示头颈部鳞癌发生、发展的多阶段演进过程.     

喉癌端粒酶活性检测及其与颈淋巴结转移的相关性

目的 :探讨喉鳞状细胞癌端粒酶活性表达及其与颈淋巴结转移的相关性。方法 :采用多聚酶链反应 -酶联免疫吸附法 (PCR- EL ISA) ,对 47例喉鳞状细胞癌组织及 10例声带炎性息肉组织的端粒酶活性进行定量检测。结果 :47例喉鳞癌组织中 39例端粒酶呈阳性 (83% ) ,而 10例声带炎性息肉组织均为阴性 ,两组差异有极显著性意义 (P <0 .0 1) ;端粒酶活性在伴有颈淋巴结转移的喉癌中显著高于无淋巴结转移者 (P <0 .0 5 )。结论 :端粒酶在喉癌组织中有较高的表达 ,可作为喉癌的肿瘤标志物。喉癌组织端粒酶活性有可能作为预测喉癌转移和预后的指标 ,并可望指导临床有针对性地应用选择性颈廓清术。     

头颈部非鳞癌组织端粒酶活性的定量检测

研究表明 ,端粒酶在人类恶性肿瘤组织中呈高度表达 ,而在正常细胞呈低表达或无表达 ,提示端粒酶表达在恶性肿瘤的发生过程中起着重要的作用。已有报道显示 ,头颈部鳞癌中端粒酶呈普遍表达〔1～ 3〕。然而 ,除少数有关端粒酶在甲状腺腺癌中表达的报道外〔4～6〕,罕见其在头颈部非鳞癌恶性肿瘤组织中表达的报道。本研究用液体闪烁计数法〔3〕,旨在了解端粒酶在头颈部非鳞癌恶性肿瘤组织中的活性 ,探讨端粒酶活性定量检测的临床价值。1　材料与方法　　共检测 1 4例头颈部非鳞癌恶性肿瘤患者的组织样本 2 5份 (其中 1 1份取自恶性肿瘤患者的相…     

头颈部恶性肿瘤组织中端粒酶活性定量检测及其意义

目的：探讨头颈部恶性肿瘤组织中端粒酶活性水平及其与肿瘤发生、发展的关系。方法：用改进的PCR—TRAP法对36例头颈部肿瘤组织、正常组织、癌旁组织分别进行端粒酶活性定量检测。结果：36例头颈部恶性肿瘤中有32例(88．9％)阳性表达，癌旁组织有4例(11．1％)阳性表达，正常组织有l例(2．8％)阳性表达，肿瘤组织与癌旁组织和正常组织之间，差异有显著性意义(P＜0．05)。端粒酶活性水平与临床分期、病理类型和分化程度不呈线性相关，癌旁组织和正常组织阳性病例的端粒酶定量水平较肿瘤组织低。结论：端粒酶的激活与头颈部恶性肿瘤的发生、发展可能密切相关；端粒酶活性有可能成为恶性肿瘤分子诊断的标志物和治疗的新靶点。     

声门上型喉癌中端粒酶活性的检测及其意义

目的：探讨端粒酶活性在声门上型喉癌中的表达及其意义。方法：采用端粒重复序列扩增法检测了ＨＥＰ－２喉癌细胞系,２６例声门上型喉癌组织和１５例癌旁组织。结果：ＨＥＰ－２喉癌细胞系呈端粒酶阳性,癌组织的端粒酶阳性率为８４．６％,明显高于癌旁组织的阳性率４０％（Ｐ〈０．０１）,喉癌组织端粒酶活性的表达与临床分期和有无颈淋巴结转移未出现出相关性（Ｐ〉０．０５）。结论：端粒酶活化是喉癌发生的重要遗传学改变,但     

头颈部鳞癌颈淋巴结隐匿性转移

目的 探讨头颈部鳞癌隐匿性颈淋巴结转移的特点和规律。方法 对111例头颈部鳞癌N_0M_0患者的颈淋巴结清扫标本进行切片观察。结果 隐匿性转移总体发生率为26.12％(29/111)。其中口腔癌18.75％(15/80),口咽癌25.00％(1/4),下咽癌54.54％(6/11),喉癌43.75％(7/16)。原发癌临床分期、肿瘤细胞分化程度是影响颈淋巴结隐匿性转移的重要因素。111例N_0M_0患者5年生存率为66.7％,其中pN~-为74.39％(61/82),pN~ 为44.82％(13/29)。结论 对临床T_3和T_4期、癌组织分化程度低和深度浸润的cN_0头颈部鳞癌应行选择性颈清扫术以治疗颈淋巴结隐匿性转移并提高患者的生存率。     

头颈部鳞癌颈淋巴结隐匿性转移

目的探讨头颈部鳞癌隐匿性颈淋巴结转移的特点和规律。方法对111例头颈部鳞癌N0M0患者的颈淋巴结清扫标本进行切片观察。结果隐匿性转移总体发生率为26．12％(29／111)。其中口腔癌18．75％(15／80)，口咽癌25．00％(1／4)，下咽癌54．54％(6／11)，喉癌43．75％(7／16)。原发癌临床分期、肿瘤细胞分化程度是影响颈淋巴结隐匿性转移的重要因素。111例N0M0患者5年生存率为66．7％，其中pN^-为74．39％(61／82)，pN^ 为44．82％(13／29)。结论对临床T3和T4期、癌组织分化程度低和深度浸润的cN0头颈部鳞癌应行选择性颈清扫术以治疗颈淋巴结隐匿性转移并提高患者的生存率。     

喉癌和癌旁组织的端粒酶活性检测

目的：探索端粒酶活性与喉癌发生发展的关系。方法：采用重复序列扩增法（TRAP－PCR）检测56例手术切除的喉癌组织和癌旁粘膜组织的端粒酶活性。喉癌组织均经病理证实,喉癌旁粘膜组织中有正常喉粘膜41例,轻度不典型增生15例。结果：喉癌组织端粒酶活性阳性率为91．07％（51／56）,正常喉粘膜和轻度不典型增生喉粘膜的阳性率分别为9．76％（4／41）和33．33％（5／15）,喉癌组织和癌旁粘膜组织中端粒酶活性阳性率有显著差异（P<0．01）。癌旁上皮端粒酶活性阳性的9例患者其喉癌组织端粒酶活性皆为阳性。结论：端粒酶激活与喉癌的发生发展有密切关系,并可作为喉癌分子诊断的肿瘤标记物。     

X染色体灭活方式检测在诊断头颈鳞状细胞癌颈淋巴微转移中的作用

目的探讨X染色体灭活检测肿瘤细胞克隆来源在诊断头颈鳞癌颈淋巴结微转移中的作用。方法对20例临床N0M0头颈部鳞癌包括10例术后病理诊断明确的颈淋巴结微转移癌和10例可疑颈淋巴结微转移癌，通过组织显微切割和蛋白酶K消化技术获得肿瘤组织DNA，在限制性酶切和PCR扩增后观察肿瘤细胞雄激素受体在X染色体上的标志，明确X染色体灭活情况；通过比较原发癌和转移灶或可疑微转移灶细胞的检测结果鉴定转移部位细胞与原发癌肿瘤细胞的克隆同源性，进而对颈淋巴转移情况做出正确诊断。结果10例不同程度表达肿瘤细胞表面标志、病理诊断明确的颈淋巴结转移癌与其对应的原发癌均为单克隆来源，并且具有相同的X染色体灭活方式，提示两者之间具有相同的克隆来源，证明颈淋巴结转移癌来自原发癌；10例原发灶不同程度地表达表皮生长因子受体（epidermal growth factor receptor，EGFR）和角蛋白，但颈淋巴结内可疑微转移灶不表达EGFR和角蛋白的可疑颈淋巴结微转移癌，X染色体灭活分析发现6例可疑转移灶细胞与原发癌细胞具有克隆同源性，证实为颈淋巴结内微转移；其余4例两者间不具有相同的细胞克隆来源，排除淋巴结转移癌。结论采用x染色体灭活法检测肿瘤细胞的克隆来源对诊断头颈肿瘤颈淋巴结微转移具有很好的应用前景和潜在临床实用价值。     

Quantitative assay of telomerase activity in head and neck squamous cell carcinoma and other tissues

OBJECTIVES: To confirm the applicability and use of a new technique to detect and quantify telomerase activity of specimens from head and neck malignant neoplasms and to explore whether the levels of telomerase activity can be a useful marker for cancer risk assessment in head and neck malignant neoplasms. DESIGN: Ninety-six specimens from 39 patients with head and neck malignant neoplasms were obtained. The specimens included 39 from patients with primary tumors (25 with head and neck squamous cell carcinoma and 14 with others), 10 from patients with neck metastases, 10 from patients with dysplasias, and 37 from patients with normal tissue. HeLa cell lines were used as positive control samples. MAIN OUTCOME MEASURE: The levels of telomerase activity were determined using a liquid scintillation counter. RESULTS: The new method has a high rate of outcome reproducibility. The intrabatch and extrabatch variations were 15.6% and 16.4%, respectively. The linear relationship was good between the telomerase activity and the value within 700 radioactive cpm (rcpm) to approximately 7000 rcpm. The levels of telomerase activity determined by radioactive count were more than 1000 rcpm in 42 of the 49 malignant specimens and much more than that in the normal tissues, with the exception of 3 specimens. The levels of telomerase activity in normal tissues were less than 1000 rcpm in every sample and less than that in the malignant neoplasm samples, with the exception of 1 specimen (P < .000). Higher levels of telomerase activity in 2 of 10 tissues from patients who had dysplasias were detected (2 specimens from patients who had severe dysplasia). The differences in the levels of telomerase activity between the head and neck squamous cell carcinomas and the other tumors were not statistically significant (P > .05). CONCLUSIONS: Detection of telomerase activity in head and neck malignant neoplasms can be a useful marker for the assessment of cancer. Telomerase reactivation may play an important role in tumorigenesis in head and neck squamous cell carcinoma. The quantification of telomerase activity may have clinical diagnostic value for head and neck malignant neoplasms.     

Expression of EphB4 in head and neck squamous cell carcinoma

EphB4 is a receptor tyrosine kinase that is expressed on epithelial cells during fetal life. It is also expressed on some venous endothelial cells. We conducted a study of six men with primary squamous cell carcinoma of the head and neck (HNSCC) that had metastasized to the cervical lymph nodes. Our goal was to determine if EphB4 is aberrantly expressed in cases of HNSCC and to determine if there is a qualitative difference between the expression of EphB4 on primary and metastatic tumors and its expression on normal mucosa adjacent to primary tumors. From each patient, we obtained specimens of the primary tumor, the nodal metastasis, and the adjacent normal mucosa, and we performed immunocytochemistry on each. We observed EphB4 expression in all primary and metastatic tumors and no expression in the normal tissue. In each of the six patients, expression was greater in the metastatic tumor than in the primary tumor. We conclude that EphB4 is a novel target in the treatment of HNSCC.     

Expression of VEGF, HGF, IL-6, IL-8, MMP-9, Telomerase in Peripheral Blood of Patients with Head and Neck Squamous Cell Carcinoma

Objectives

This study investigated the telomerase expression in peripheral blood mononuclear cells (PBMCs) and the relationship between the serum level of several soluble factors such as vascular endothelial growth factor (VEGF), hepatocyte growth factor, interleukin (IL)-6, IL-8, and matrix metallopeptidase-9 and the clinicopathological features of patients with head and neck squamous cell carcinoma (HNSCC).

Methods

Peripheral blood samples were collected from 50 HNSCC patients and 15 normal controls. The telomerase activity in the PBMCs was measured by Telomere Repeat Amplification Protocols. The serum levels of the soluble factors were analyzed by enzyme-linked immunosorbent assay.

Results

The expression of telomerase in the PBMCs of HNSCC patients was significantly correlated with the N and American Joint Committee on Cancer (AJCC) stages. The serum VEGF level was significantly higher in the patients with an advanced T stage, N stage and AJCC stage. Serum VEGF was significantly related with the expression of telomerase in the PBMCs. The telomerase expression and the VEGF expression were shown to be independent factors associated with poor survival.

Conclusion

The telomerase expression in the PBMCs and the serum VEGF level of HNSCC patients were significantly correlated with the N stage, the AJCC stage and the prognosis.     

Expression der immun- supprimierenden Zytokine TGF-β2 und Il-10 in Kopf-Hals-Tumoren Vergleich von Serumwerten und Gewebeexpression

Transforming growth factor-beta (TGF-beta) and interleukin 10 (Il-10) are cytokines that have a strong immunosuppressive ability. Their secretion by tumor cells is able to suppress an immunological response against tumor. Both factors have been shown to enhance tumor growth in glioblastomas and carcinoma of the breast. We determined the expression pattern of TGF-beta and Il-10 in squamous cell carcinomas of the head and neck (HNSCC) and a possible association with tumor stage and their pre-treatment cytokine serum levels. Cytokine expression in primary tumors and metastases of 21 patients with HNSCC was investigated by immunohistochemistry. To assess the TGF-beta2 and Il-10 levels in tumor patients before therapy 49 serum specimens were analyzed by ELISA. TGF-beta2 was detected in 95% of all tumor tissues analyzed and Il-10 in 79% of all tumors. TGF-beta2 was localized in tumor cells and tumor borders, while Il-10 was preferentially found in peritumoral connective tissue. Metastasizing tumors showed elevated pretreatment serum levels for TGF-beta2 and Il-10. There was no correlation between TGF-beta2 and Il-10 expression in tumor tissue and pretreatment serum levels. Our data show that the majority of HNSCC analyzed express TGF-beta2 and Il-10. A correlation between pretherapy elevated cytokine serum levels and tumor grade was shown.     

Zwint-1蛋白在头颈部鳞状细胞癌中表达上调及其与临床特征的相关性

目的探讨Zeste white 10（ZW10）相互作用着丝粒蛋白1 (ZW10 interacting kinetochore protein 1,Zwint 1),在头颈部鳞状细胞癌（head and neck squamous cell carcinoma,HNSCC）肿瘤组织中的蛋白表达情况,以及其与临床病理特征之间的关系。方法采用免疫组织化学检测的方法,对16例HNSCC患者的肿瘤组织样本,以及7例相应的癌旁正常组织样本中Zwint 1蛋白进行检测。同时,分析HNSCC相关组织芯片中Zwint 1蛋白表达与HNSCC临床特征的相关性。结果临床样本与组织芯片的检测结果显示,Zwint 1蛋白在HNSCC肿瘤组织中的蛋白表达高于正常组织,其差异具有统计学意义（t=2.399, P<0.05）。组织芯片的检测结果显示,Zwint 1蛋白表达量与HNSCC患者的肿瘤大小（F=2.889,P<0.05）、淋巴结转移（t=2.110,P<0.05）、分化（F=3.667,P<0.05）以及临床分期（F= 2.864,P<0.05）均具有相关性。但是卡方检验结果显示,Zwint 1蛋白阳性表达与肿瘤大小（χ2=3.236,P>0.05）、淋巴结转移（χ2=1.463,P>0.05）、分化（χ2=1.271,P>0.05）、临床分期（χ2=4.179,P>0.05）,差异无统计学意义。结论Zwint 1蛋白在HNSCC的发生发展过程中发挥有重要作用,可能是HNSCC潜在的靶向治疗目标。     

Clinical and molecular characteristics of HNSCC patients with brain metastases: a retrospective study

Among the metastasis patterns of head and neck squamous cell carcinoma (HNSCC), intracranial spread is a rare but dreaded event. To date only very few cases have been reported and clinical and molecular data are sparse. We screened our archives for HNSCC patients from 1992 to 2005 who were diagnosed with brain metastases (BM). For retrospective analysis, all clinico-pathological data including disease-free survival (DFS), local progression-free survival (LPFS), and overall survival (OS) were compiled. Additionally, we assessed the mutational status of the TP53 gene and the prevalence of HPV serotypes by PCR and Sanger sequencing. Immunohistochemistry was applied to detect p16INK4A expression levels as surrogate marker for HPV infection. The prevalence rate of BM in our cohort comprising 193 patients with advanced HNSCC was 5.7 %. Of 11 patients with BM, 3 were female and 9 were male. Seven of the primary tumors were of oropharyngeal origin (OPSCC). LPFS of the cohort was 11.8 months, DFS was 12.1 months and OS was 36.0 months. After the diagnosis of BM, survival was 10.5 months. Five tumors showed a mutation in the TP53 gene, while five of the seven OPSCC tumors had a positive HPV status displaying infection with serotype 16 in all cases. Compared with patients who harbored TP53wt/HPV-positive tumors, patients with TP53 mutations showed a poor prognosis. Compared with the whole cohort, the interval between diagnosis of the primary and the detection of BM was prolonged in the HPV-infected OPSCC subgroup (26.4 vs. 45.6 months). The prognosis of HNSCC patients with BM is poor. In our cohort, most tumors were OPSCC with the majority being HPV positive. Our study points toward a putatively unusual metastatic behavior of HPV-positive OPSCC.     

Telomerase activity and cell proliferation index in cholesteatoma

CONCLUSIONS: Telomerase activity was expressed in cholesteatomas, and cellular proliferation was significantly higher in cases where the telomerase activity was positive. Telomerase activity was also closely related with cellular proliferation in chronic hyperproliferating tissues such as cholesteatomas. OBJECTIVE: Telomerase activity is detected in most malignant tumors and is also known to have a close relationship with cell proliferation. Cholesteatoma shows cellular hyperproliferation. We studied telomerase activity in cholesteatoma and its relationship with cellular proliferation and clinical findings. MATERIAL AND METHODS: Cholesteatoma tissue was obtained from 40 patients during middle ear surgery. Telomerase activity was measured using a telomeric repeat amplification protocol method. As a cellular proliferation index, expression of Ki-67 was measured by means of immunohistochemical staining. Posterior auricular skin was used as a control. Telomerase activity was compared with Ki-67 expression. Clinical features such as hearing loss, the extension of cholesteatoma, the degree of bone destruction and the cause of cholesteatomas were compared with telomerase activity and the cellular proliferation index. RESULTS: Telomerase activity was positive in 21/40 cholesteatomas (52.5%), but absent in the control group. The average Ki-67 labeling index in the cholesteatoma group was 32.84+/-10.13, higher than that in the control group (21.83+/-7.76) (p<0.05). The average Ki-67 labeling indices of the 21 telomerase activity-positive and 19 telomerase activity-negative cholesteatomas were 37.76+/-8.53 and 27.39+/-9.06, respectively. The Ki-67 labeling index was significantly higher in telomerase-positive cholesteatomas (p<0.05). The clinical features did not show a relationship with either telomerase activity or the cellular proliferation index.