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.

卵巢癌被认为是覆盖人卵巢自由表面的卵巢表面上皮（OSE）的细胞。先前从生殖时代患者的正常OSE细胞中建立了两种永生的人类细胞系OSE2A（非甲状腺菌）和OSE2B-2（肿瘤）。在本项目中，我们发现在OSE2A细胞中存在黄体生成马酮（LH）/人绒毛膜促性腺激素（HCG）受体（R），而OSE2B-2细胞中不存在。在OSE2A细胞中，HCG的低浓度（10^3 MIU/mL）通过上调胰岛素样生长因子1（IGF1）增强了锚固依赖性生长，而HCG诱导的锚固型独立性独立的生长和下降的HCG浓度（10^5 mLU/mL）。为了研究其他基因参与LH/HCGR相关的肿瘤性，我们比较了OSE2A和OSE2B-2细胞之间的cDNA表达阵列，发现以下基因在OSE2B-2中的表达低于OSE2A：整合蛋白β1，cellibularβ1，celllular粘贴分子 - 粘合分子-1（ICAM1）和WAFL/CILL。随后使用OSE2A细胞的半定量逆转录聚合酶链反应表明，整联蛋白β1的表达被HCG的高浓度（10^5 mlu/ml）下调。这些结果表明，LH/CGR通过介导IGF1和整联蛋白β1的上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会导致其信号转导途径的遗传改变，从而导致OSE细胞生长的刺激，卵巢致癌作用和癌症进展。我们已将本研究的手稿提交给“癌症科学。我们仍然进一步研究了PTEN表达的项目。