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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卵巢表面上皮（OSE）细胞覆盖的人卵巢的自由表面。两个永生化的人细胞系，OSE 2a（非致瘤性）和OSE 2b-2（致瘤性），以前建立从正常的OSE细胞的育龄患者。在本项目中，我们发现促黄体生成素（LH）/人绒毛膜促性腺激素（hCG）受体（R）在OSE 2a细胞中表达，而在OSE 2b-2细胞中不表达。在OSE 2a细胞中，低浓度（10^3 mIU/ml）的hCG通过上调胰岛素样生长因子-1（IGF 1）来增强锚定依赖性生长，而高浓度（10^5 mIU/ml）的hCG诱导锚定非依赖性生长和下调IGF 1表达。为了研究LH/hCG相关致瘤性中其他基因的参与，我们比较了OSE 2a和OSE 2b-2细胞之间的cDNA表达阵列，发现以下基因在OSE 2b-2中的表达低于OSE 2a：整合素β1，细胞间粘附分子1（ICAM 1）和Wafl/Cipl。随后使用OSE 2a细胞的半定量逆转录聚合酶链反应显示，整合素β1的表达被高浓度（10^5 mIU/ml）的hCG下调。这些结果表明，LH/CGR通过介导IGF 1和整合素β1的上调和下调来影响锚定依赖性和非依赖性生长。LH/hCGR的重复和过度激活可能导致其信号转导途径的遗传改变，从而刺激OSE细胞的生长，引发卵巢癌的发生和癌症的进展。我们已经向《癌症科学》提交了一份研究论文。关于PTEN基因，在研究PTEN表达载体转染永生化OSE细胞的特性之前，对LH/hCGR进行了功能分析。我们还将进一步研究PTEN的表达方案。