Detection of telomerase activity and hTERT mRNA expression and its application for the diagnosis of gynecologic tumors

端粒酶活性和hTERT mRNA表达检测及其在妇科肿瘤诊断中的应用

基本信息

批准号：
11671604
负责人：
KYO Satoru
金额：
$ 2.18万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Telomerase is a cellular reverse transcriptase which catalyzes the synthesis and extension of telomeric DNA.This enzyme is specifically activated in most malignant tumors, but is usually inactive in normal somatic cells, and within which telomeres therefore progressively shorten with cell division. A mechanism to maintain telomere stability is required for the cells to overcome replicative senescence, and telomerase activation may therefore be a rate-limiting or critical step in cellular immortality and oncogenesis. Human telomerase reverse transcriptase (hTERT) is a catalytic subunit protein of telomerase and is a critical determinant of its enzymatic activity.In the present study, we examined the utility of detecting telomerase activity and hTERT mRNA expression for the diagnosis and screening of gynecologyc tumors. We found that most gynecologic cancers express telomerase activity while the corresponding normal tissues do not. Cervical and endometrial exfoliated cells were collected from the patients with or without cervical or endometrial lesions and telomerase activity and hTERT mRNA expression were detected by the TRAP assays or RT-PCR assays. Approximately 90% of the patients with cervical or endometrial cancer were positive for telomerase activity or hTERT mRNA expression, while less than 10% of those without any lesions were so. 50 to 60% of the patients with premalignant lesions, such as suqamous intraepithelial lesions (SIL) expressed telomerase activity or hTERT mRNA.These findings suggest that detection of telomerase activity or hTERT mRNA expression may be an useful tool for cancer diagnosis and screening.

端粒酶是一种细胞逆转录酶，可催化端粒 DNA 的合成和延伸。这种酶在大多数恶性肿瘤中特异性激活，但在正常体细胞中通常不活跃，因此端粒随着细胞分裂而逐渐缩短。细胞需要一种维持端粒稳定性的机制来克服复制衰老，因此端粒酶激活可能是细胞永生和肿瘤发生的限速或关键步骤。人端粒酶逆转录酶（hTERT）是端粒酶的催化亚基蛋白，是其酶活性的关键决定因素。在本研究中，我们研究了检测端粒酶活性和hTERT mRNA表达在妇科肿瘤诊断和筛查中的实用性。我们发现大多数妇科癌症表达端粒酶活性，而相应的正常组织则不表达端粒酶活性。采集有或无宫颈或子宫内膜病变患者的宫颈和子宫内膜脱落细胞，采用TRAP法或RT-PCR法检测端粒酶活性和hTERT mRNA表达。大约 90% 的宫颈癌或子宫内膜癌患者端粒酶活性或 hTERT mRNA 表达呈阳性，而无任何病变的患者中只有不到 10% 呈阳性。 50%~60%的癌前病变，如鳞状上皮内病变（SIL）的患者表达端粒酶活性或hTERT mRNA。这些发现表明，检测端粒酶活性或hTERT mRNA表达可能是癌症诊断和筛查的有用工具。