Identification of A Group of Genes Associated with Oncogenesis of Ovarian Cancers

一组与卵巢癌发生相关的基因的鉴定

• 批准号: 08457435
• 负责人: SEKIYA Souei
• 金额: $ 5.18万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457435/
• 关键词: Ovarian cancer; Oncogenesis; Telomerase; Telomere; quantitative measurement

In this study we focused on the telomerase gene among a group of those associated with oncogenesis of ovarian cancers. By a variety of available molecular and cytogenetic techniques, cell lines and surgical specimens of ovarian cancers have been analyzed to clinically investigate following questions. 1) whether the shortening of telomere length and the acceleration of telomerase activity are correlated to carcinogenesis or not, 2) if so, to which stage in tumor progression, 3) how they communicate with other oncogenes and tumor suppresser genes.We designed new PCR primers for detecting telomerase activity, which are different from ordinary ones used in TRAP assay and succeeded in performing electrophoresis and computer analysis on an autosequencer using theses fluorescence-labeled primers. Fragment analysis program led to accurate and convenient quantitative measurement of fluorescent ladder bands produced by telomerase. This newly developed assay system was applied to quantitative ana … More lysis of telomerase activity in cell lines, primary or metastatic tumors, recurrence tumors after chemotherapy, and formalin-fixed tissue sections.In 6 ovarian cancer cell lines we examined high degrees of telomerase activity were detected as expected. However, we failed to obtain stable results for surgical and pathological specimen of ovarian cancers. This failure was thought to result from some inhibitory substance for telomerase or PCR reaction in tissue extracts. Several extraction techniques for tissue proteins were thus examined, but this problem has not been overcome in the research period. We also developed in situ detection of telomerase activity in smear cells of ovarian and cervical cancers, and obtain preliminary results promising clinical application.It is scheduled to make continuous improvements to establish quantitative measurement of telomerase activity in povarian cancers and, compare degrees of their activity with clinical background including stage, tissue type and grade, and expression of a panel of oncogenes and cell cycle-related genes. Our final goal is to clarify the roles of telomerase in oncogenesis mechanism and examine the ossibility of its clinical application. Less

在这项研究中，我们集中在端粒酶基因中的一组与卵巢癌的肿瘤发生。通过各种可用的分子和细胞遗传学技术，卵巢癌的细胞系和手术标本进行了分析，以临床研究以下问题。1)端粒长度的缩短和端粒酶活性的增强是否与肿瘤的发生有关，2）如果是，与肿瘤的哪个阶段有关，3）它们如何与其他癌基因和抑癌基因通讯。我们设计了新的PCR引物检测端粒酶活性，其不同于TRAP测定中使用的普通方法，并成功地使用这些荧光-标记引物。片段分析程序导致端粒酶产生的荧光梯状条带的准确和方便的定量测量。将该系统应用于定量分析， ...更多信息 在细胞系、原发性或转移性肿瘤、化疗后复发的肿瘤和福尔马林固定的组织切片中的端粒酶活性的裂解。在我们检测的6个卵巢癌细胞系中，如预期的那样检测到高度端粒酶活性。然而，我们未能获得稳定的结果，手术和病理标本的卵巢癌。这种失败被认为是由于组织提取物中的某些端粒酶抑制物质或PCR反应。因此，研究了几种组织蛋白的提取技术，但在研究期间，这个问题尚未得到解决。我们还开展了卵巢癌和宫颈癌涂片细胞端粒酶活性的原位检测，取得了初步的临床应用效果，并计划继续改进，建立卵巢癌端粒酶活性的定量检测方法，并将其活性程度与临床背景（包括分期、组织类型和分级）进行比较，以及一组癌基因和细胞周期相关基因的表达。我们的最终目的是阐明端粒酶在肿瘤发生机制中的作用，并探讨其临床应用的可能性。少