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     

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。     

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。     

凋亡抑制因子Survivin在喉癌中的表达及其意义

检测凋亡抑制因子Survivin在喉癌组织中的表达及其与喉癌的临床、病理特性之间的关系。方法：取石蜡包埋喉鳞状细胞癌标本33例，声带息肉21例，应用免疫组化SABC法检测Survivin在喉癌和声带息肉组织中的表达。结果：Survivin在喉癌组织中的表达率为63.6%,在声带息肉组织中的表达率为28.6%,二者有统计学差异（P<0.05）。而Survivin的表达与喉癌的临床分期、组织学分级、淋巴结转移和患者年龄、性别等无关。结论：凋亡抑制因子Survivin在喉癌组织中高表达，在声带息肉中低表达，与喉癌的发生、发展密切相关。     

人端粒保护蛋白POT1和TRF2在喉癌中的表达

目的探讨人端粒保护蛋白1（human protection of telomeres1,POT1）和端粒重复序列结合因子2（telomeric repeat binding factor-2,TRF2）在喉鳞状细胞癌（简称喉癌）及声带息肉中的表达及意义。方法采用间接免疫荧光法检测20例喉癌、19例声带息肉中POT1和TRF2的表达水平。结果POT1和TRF2在喉癌中的阳性表达率为65．00％和70．00％,在声带息肉中未见POT1和TRF2的表达。POT1在低分化喉癌中的表达高于POT1在高及中分化喉癌中的表达（P〈0．05）,POT1的表达水平在喉癌的不同发生部位、T分级、临床分期及淋巴结转移率中的差异无统计学意义（P〈0．05）。TRF2的表达程度与肿瘤的发生部位、组织病理、T分级、淋巴结转移率和临床分期均无关（P〈0．05）。相关分析显示喉癌中POT1的表达与TRF2的表达无相关性（P〈0．05）。结论POT1和TRF2在喉癌中高表达,两者在喉癌的发生中可能起重要作用。POT1在喉癌中的表达与肿瘤的分化程度有关。     

喉鳞癌端粒酶及其端粒酶RNA的表达

目的：探讨端粒酶及其组成人端粒酶ＲＮＡ（ｈＴＲ）在喉鳞癌中的表达。方法：分别采用ＴＲＡＰ ＰＣＲ ＰＡＧＥ凝胶电泳及巢式ＲＴ－ＰＣＲ测定５例喉鳞癌及３例对照粘膜中的端粒酶及ｈＴＲ。结果：５例喉鳞癌端粒酶表达均为阳笥，３例对照组则均为阴必。所有检测标本的ｈＴＲ均为阳性。结论：端粒酶在喉鳞癌呈现高表达，而正常喉粘膜则无。ｈＴＲ广泛存在于喉癌与非肿瘤组织中，因而仅从ｈＴＲ的表达不能说明与喉部高分化鳞癌的     

免疫组化法检测喉癌组织中p53表达的临床意义

采用免疫组化Ｓ－Ｐ方法检测９５例喉癌、１０例声带息肉和６例喉正常声带组织的ｐ５３表达，结果显示，ｐ５３蛋白主要定位于喉癌细胞内，呈棕黄色阳性颗粒，少部分位于细胞浆内，９５例中阳性显色４２例，阳性率４４．２％。１０例喉气带息肉和６例喉正常声带组织均为阴性。喉癌中ｐ５３基因表达阳性与声带息肉、正常声带组织相比差异显著（Ｐ〈０．０１）。喉癌组中，不同年龄组发病有显著性差异（Ｐ〈０．０５）。喉癌不同临床分     

喉癌标本端粒酶活性的检测

目的：探讨端粒酶生在喉癌发生过程中的作用。方法：采用ＴＲＡＰ方法检测３４例头颈肿瘤组织标本中的端粒酶活性。结果：在２７例喉癌患者中有２３例同端粒是笥，阳性率８５．２％，２７例相应癌旁组织有７例检出端粒酶阳性，阳性率２５．９％，７例喉乳头瘤瘤患者中有３例检出，阳性率４２．９％，结论：端粒酶活化并非只发生在喉癌进展的晚期阶段，在肿瘤形成的早期也有一定程度的端粒酶激活。端粒酶活性可能民癌的恶性程度有关。     

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     

转化生长因子β_1蛋白在原发性喉癌及喉旁组织和声带息肉中表达的研究

为探讨转化生长因子β1在原发性喉癌中的表达情况，采用免疫组化LSAB方法对36例原发性喉癌及其边缘1．0cm组织和20例同年龄段声带息肉组织进行检测，喉癌组阳性车38．9％，癌旁组织组66．7％，声带息肉组60．0％，喉癌组与癌旁组织组阳性率差异显著（P＜0．05），喉癌组及声带息肉组中阳性与强阳件表达率差异显著（均P＜0．01），癌细胞分化越高TGF-β1表达越高。提示TGF-β1对喉癌细胞的增殖调控起着重要作用。     

喉鳞癌端粒酶活性与细胞周期蛋白D1的表达及其意义

目的 :了解喉鳞癌端粒酶活性的表达情况及其与细胞周期蛋白D1(cyclinD1)过度表达的关系。方法 :分别应用TRAP法和免疫组化法对 38例喉鳞癌组织及相关喉粘膜组织的端粒酶活性和cyclinD1的表达情况进行检测。结果 :喉鳞癌病例中 82 % (31/ 38)有端粒酶活性表达 ,5 0 % (19/ 38)有cyclinD1过度表达 ;两者表达均与患者的年龄、肿瘤部位、T分期、病理学分级无关 (P >0 .0 5 ) ,而端粒酶表达与肿瘤的临床分期有关 (P <0 .0 5 ) ,cyclinD1表达与肿瘤的临床分期无关 (P >0 .0 5 ) ,喉正常粘膜中两者均无表达 ;喉鳞癌组织中两者均表达 18例 ,两者密切相关 (P <0 .0 5 )。结论 :端粒酶的激活在喉鳞癌组织中普遍存在 ,cyclinD1过度表达可能是喉鳞癌端粒酶激活的重要机制之一。     

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.     

丁酸钠对Hep-2细胞的生长抑制和诱导凋亡作用及其对端粒酶活性的影响

目的 研究丁酸钠对Hep-2细胞的生长抑制和诱导凋亡作用及诱导凋亡过程中对端粒酶活性的影响.方法 采用四甲基偶氮唑蓝(methyl thiazolyl terazolium,MTT)比色法观察不同浓度丁酸钠对Hep-2细胞的生长抑制作用,透射电镜下观察其形态学变化,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling,TUNEL)法、琼脂糖凝胶电泳和流式细胞仪检测丁酸钠作用后的细胞凋亡情况及细胞周期变化,端粒重复序列扩增(telomeric repeat amplification protocal,TRAP)-银染法检测端粒酶活性变化,RT-PCR分析端粒酶各组分的mRNA表达情况.结果 丁酸钠对Hep-2细胞的生长抑制作用呈时间和剂量依赖性.透射电镜下可见典型的凋亡细胞形态学改变.琼脂糖凝胶电泳可见特征性的凋亡细胞DNA梯状条带.TUNEL法显示,2.5 mmol/L丁酸钠作用72 h,细胞凋亡指数由作用前的2.27±1.18增加至33.50±2.75.流式细胞仪显示,2.5 mmol/L丁酸钠作用72 h,细胞凋亡率由作用前的2.86%增加至31.28%,G0/G1期细胞比例由50.38%增加至70.88%,S期细胞比例由27.40%减少至8.20%,细胞增殖指数由49.62%降低至29.12%.TRAP-银染法显示,丁酸钠作用后细胞端粒酶活性下降,且具有一定时间效应关系.RT-PCR显示端粒酶逆转录酶表达下降而端粒酶RNA模板和端粒酶相关蛋白表达无明显改变.结论 丁酸钠对Hep-2细胞具有生长抑制和诱导凋亡作用,并可能通过下调端粒酶逆转录酶表达而抑制端粒酶活性.     

癌旁喉黏膜端粒酶活化的意义

目的 :研究癌旁非癌变喉黏膜端粒酶的表达 ,探讨端粒酶的活化在喉癌扩展及复发中的作用。方法 :采用端粒重复序列扩增法 ,结合组织病理学观察。结果 :癌旁阴性切缘以外喉黏膜的端粒酶阳性率高于喉癌组织 (P <0 .0 5 )。结论 :端粒酶的激活发生在喉癌的形态学改变之前 ,可能提示有恶性转化潜能的细胞扩散。     

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.     

Expression patterns of Ki-67 and telomerase activity in middle ear cholesteatoma.

BACKGROUND: Keratinocytes in cholesteatoma demonstrate uncoordinated hyperproliferation, migration, and invasion properties. There is a controversy regarding the impact of Ki-67 and telomerase activities on cellular proliferation in cholesteatoma. We studied expression of Ki-67 protein and telomerase activity in cholesteatoma and its relationship with clinical findings. METHODS: The expression level of Ki-67 protein was examined by immunohistochemical analysis of 51 cholesteatomas and 6 skin tissues obtained from patients during ear surgery. Telomerase activity was determined in 23 samples of cholesteatomas and 6 skin samples by polymerase chain reaction-based telomeric-repeat amplification protocol assay. RESULTS: The presence of Ki-67 protein was observed in 21 (41.2%) of 51 samples of acquired cholesteatoma. The average Ki-67 labeling index in the cholesteatoma group was 28.9 +/- 9.2 and was higher than that in the skin group (18.2 +/- 6.1). Telomerase activity was detected in 2 (8.7%) of 23 samples of cholesteatoma (21 of them were Ki-67 staining positive and 2, negative) and in 3 (50%) of 6 of control skin samples (p < 0.05). CONCLUSION: This study showed increased expression of Ki-67 in cholesteatoma, whereas there was no significant difference in rate of Ki-67 positive staining between skin and cholesteatoma (p = 0.066). Telomerase activation is a rare event in cholesteatoma. We assume that the absence of telomerase may lead to generation dysfunctional telomeres what in turn may impair the proliferative capacity of cholesteatoma.