支气管肺泡灌洗液脱落细胞端粒酶hTERT基因表达的检测

背景与目的:肺癌患者中端粒酶活性常呈阳性,常规检测端粒酶活性的方法易发生假阳性,端粒酶hTERT基因表达水平可反映端粒酶活性高低。本研究采用相对定量PT-PCR法检测肺癌支气管肺泡灌洗液脱落细胞端粒酶hTERT的表达情况,为肺癌的早期诊断、鉴别诊断提供帮助。方法:采用纤维支气管镜收集了20例可疑肺肿瘤患者支气管肺泡灌洗液,对脱落细胞分别送病理检测和RT-PCR相对定量法检测端粒酶hTERT基因表达,并随访术后病理检测情况,同时检测10例非肺癌病人的支气管肺泡灌洗液标本作为对照。结果:20例可疑肺癌经术后病理检测证实有19例为不同类型的肺癌,1例为炎性假瘤。在19例肺癌的支气管肺泡灌洗液中16例检出不同程度的端粒酶hTERT基因表达,细胞病理检测仅发现11例,阳性率分别为84.2%和57.9%,两者有显著性差异(P<0.01)。10例非肺癌病人的支气管肺泡灌洗液中均无端粒酶hTERT基因表达。结论:采用RT-PCR方法检测支气管肺泡灌洗液中端粒酶hTERT基因表达,较细胞病理检测有更高的敏感性,有助于肺癌的早期诊断和鉴别诊断。     

Differential telomerase activity, expression of the telomerase catalytic sub-unit and telomerase-RNA in ovarian tumors.

Telomerase activity has been found in a variety of malignant tumors but only rarely in benign tumors or normal tissues. In this study, we investigated telomerase activation in 37 ovarian tumors, including benign, borderline and malignant neoplasms. Telomerase activity was detected using the telomeric repeat amplification protocol (TRAP) in 13/16 ovarian carcinomas, 9/10 borderline tumors and 3/11 cystadenomas/fibromas. mRNA expression of the putative human telomerase catalytic sub-unit gene (hTERT) was detected by RT-PCR in 14/15 ovarian carcinomas, 8/10 borderline tumors and 4/11 cystadenomas/fibromas. In situ hybridization was performed to evaluate telomerase-RNA (hTR) expression in the corresponding paraffin-embedded tumors. Variable expression levels of hTR were found over neoplastic tumor cells. The highest levels of hTR expression were found predominantly in ovarian carcinomas. Although the amount of telomerase activity varied, significantly high levels of telomerase activity were found predominantly in ovarian carcinomas. hTERT mRNA expression was closely associated with telomerase activity. These findings suggest that up-regulation of hTERT and hTR is important for telomerase activation during malignant-tumor progression. Telomerase activation might therefore be a valuable diagnostic parameter that could help to identify potentially progressive lesions. However, the diagnostic and therapeutic implications of telomerase activation need to be clarified in clinical trials. Int. J. Cancer (Pred. Oncol.) 84:426-431, 1999.     

宫颈癌及子宫内膜癌细胞系中hTERT启动子活性的研究

目的 :探讨Hela及HHUC细胞系中hTERT启动子的活性及其与hTERTmRNA表达和端粒酶活性之间的关系。方法 :将hTERT核心启动子基因转入Hela、HHUC细胞系、人正常皮肤表皮细胞及子宫内膜上皮细胞中 ,检测细胞系hTERT启动子的活性、hTERTmRNA表达水平及端粒酶活性。分别以HEK2 93细胞系作为阳性对照 ,HELF细胞系作为阴性对照。结果 :细胞系Hela、HHUC、人正常皮肤表皮细胞及正常子宫内膜上皮细胞中的hTERT启动子活性分别为 2 0 1%、14 9%、0 3%和 0 5 % ;hTERTmRNA相对表达水平分别为 0 6 3、0 5 1、0和 0 ;端粒酶活性分别为 0 393、0 387、0 14 4和 0 15 2。结论 :宫颈癌细胞系及子宫内膜癌细胞系中hTERT启动子活性、hTERTmRNA表达水平和端粒酶活性均特异性增高 ,而在正常角化细胞及子宫内膜上皮细胞中则被抑制或不表达。hTERT启动子活性水平与hTERTmRNA表达水平和端粒酶活性之间有显著相关性