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Characterization of immortalized human ovarian surface epithelial cells transfected by PTEN expression vectors

PTEN表达载体转染的永生化人卵巢表面上皮细胞的表征

基本信息

批准号：
12671616
负责人：
KATABUCHI Hidetaka
金额：
$ 2.11万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671616/
关键词：
ovarian surface epithelium epithelial ovarian carcinoma PTEN LH hCG receptor Cip1 Waf1 ICAM-1 integrin anchorage dependent and -independent growth PTEN遺伝子 LH受容体遺伝子 Waf1遺伝子 ICAM1遺伝子足場非依存性増殖 Cip2遺伝子造腫瘍性

项目摘要

Epithelial ovarian carcinomas are thought to arise from cells of ovarian surface epithelium (OSE) covering the free surface of the human ovary. Two immortalized human cell lines, OSE2a (non-tumorigenic) and OSE2b-2 (tumorigenic), were previously established from normal OSE cells of a reproductive-age patient. In the present project, we found that expression of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor (R) is present in OSE2a cells and absent in OSE2b-2 cells. In OSE2a cells, a low concentration (10^3 mIU/ml) of hCG enhanced anchorage-dependent growth via up-regulation of insulin-like growth factor-1 (IGF1), whereas a high concentration (10^5 mlU/ml) of hCG induced anchorage-independent growth and down-regulation of IGF1 expression. To investigate involvement of other genes in LH/hCGR-related tumorigenicity, we compared cDNA expression arrays between OSE2a and OSE2b-2 cells, and found that the following genes had lower expression in OSE2b-2 than in OSE2a: integrin β1, intercellular adhesion molecule-1 (ICAM1), and Wafl/Cipl. Subsequent semiquantitative reverse transcription porymerase chain reaction using OSE2a cells showed that expression of integrin β1 was down-regulated by a high concentration (10^5 mlU/ml) of hCG. These results suggest that LH/CGR affects anchorage-dependent and -independent growth by mediating up- and down-regulation of IGF1 and integrin β1. Repetitive and excessive activation of LH/hCGR may cause genetic alteration of its signal transduction pathway, resulting in stimulation of growth of OSE cells, initiation of ovarian carcinogenesis, and cancer progression. We have submitted a manuscript for this study to 'Cancer Science'.Regarding the PTEN gene, the functional analysis of LH/hCGR preceded the study for characterization of immortalized OSE cells trasfected by PTEN expression vectors. We still farther study the project for the PTEN expression.

上皮性卵巢癌被认为是由覆盖在人卵巢游离表面的卵巢表面上皮(OSE)细胞引起的。两个永生化的人类细胞系，OSE2a(非致癌)和OSE2b-2(致癌)，以前是从一名生殖年龄患者的正常OSE细胞建立的。在本课题中，我们发现促黄体生成素/人绒毛膜促性腺激素受体(R)在OSE2a细胞中表达，而在OSE2b-2细胞中不表达。在OSE2a细胞中，低浓度(10.3mIU/ml)hCG通过上调胰岛素样生长因子-1(IGF1)促进锚定依赖性生长，而高浓度(10.5mLU/ml)hCG则诱导非锚定生长和IGF1表达下调。为了研究其他基因在促黄体生成素/人绒毛膜促性腺激素受体相关肿瘤发生中的作用，我们比较了OSE2a和OSE2b-2细胞的基因表达谱，发现以下基因在OSE2b-2中的表达低于在OSE2a中的表达：整合素β1、细胞间黏附分子1和WAFL/Cip1。OSE2a细胞半定量逆转录聚合酶链式反应显示，高浓度(105mLU/ml)的hCG可下调整合素β-1的表达。这些结果提示，促黄体生成素/促性腺激素受体通过上调和下调胰岛素样生长因子1和整合素β-1来影响锚定依赖性和非依赖性生长。重复和过度激活促黄体生成素/促性腺激素受体信号转导通路可能导致其信号转导通路的基因改变，从而刺激卵巢上皮性癌细胞的生长，启动卵巢癌的发生和发展。我们已经向《癌症科学》杂志提交了这项研究的稿件。关于PTEN基因，在PTEN表达载体导入永生化OSE细胞的研究之前，对PTEN基因的功能分析先于对永生化OSE细胞的研究。我们还需要进一步研究PTEN的表达方案。