HCAP1两种基因型对肝癌细胞系Hep3B基因表达的影响  

Effects of Two Haplotypes of HCAP1 Gene on Cellular Gene Expression Levels in a Human Hepatocarcinoma Cell Line Hep3B

作　　者：王俊茹[1] 覃文新[1] 何祥火[1] 潘勇[1] 胡建[1] 万大方[1] 顾健人[1] 

机构地区：[1]上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032

出　　处：《癌症》2002年第11期1173-1181,共9页Chinese Journal of Cancer

基　　金：国家"863"生物高技术资助项目(批准号Z19-02-01-01);国家重点基础研究发展规划"973"资金资助项目(批准号G1998051209)

摘　　要：背景与目的:HCAP1(HCC-associatedprotein1)基因是我们室克隆的一个新基因,HCAP1基因有N和M两种基因型,本研究旨在探讨HCAP1N和HCAP1M基因型对肝癌细胞基因表达的影响。方法:用脂质体转染试剂,将HCAP1M与HCAP1N分别转染Hep3B-Tet-on肝细胞性肝癌细胞株;为了获得高诱导表达HCAP1N或HCAP1M的细胞株,通过Westernblot方法筛选被转染的细胞。然后,通过人cDNAarray膜片杂交技术分析HCAP1N或HCAP1M诱导表达前后的细胞基因表达谱。杂交结果用复日Smartview2001图像分析软件进行分析比较。结果:建立了HCAP1N和HCAP1M过表达前后的差异基因表达谱。结果表明HCAP1M过表达引起参与细胞增殖的基因表达上调(如c-erbB2受体和转录因子DP2),以及起凋亡作用基因(caspase9)及DNA切割修复蛋白基因(ERCC5)的表达下调;而HCAP1N过表达引起抑制细胞生长基因(如snoN和WAF1)和DNA切割修复蛋白基因(ERCC5)的表达上调,以及抗凋亡基因(HSPs)的表达下调。结论:HCAP1M基因过表达可能促进细胞过度增殖,HCAP1N过表达可能抑制细胞生长。Background and Objective: HCAP1 gene is a novel gene cloned in our laboratory, which has two haplotypes(N and M). This study was designed to investigate the effects of two haplotypes of HCAP1 gene on cellular gene expression levels in a human hepatocarcinoma cell line Hep3B. Methods: The Hep3B Tet on cells were transfected with HCAP1N or HCAP1M using liposome transfection reagent. In order to obtain the cells with high inducible expression of HCAP1N or HCAP1M, the transfected cells were screened by Western blot. Then, human atlas cDNA expression arrays were used to analyze the cellular gene expression profiles before and after HCAP1N or HCAP1M over expression. The hybridization results were analyzed with FR 200 image system. Results: The profiles of differential gene expression before and after HCAP1N or HCAP1M over expression were established. These data indicated that over expression of HCAP1M resulted in an up regulation of genes involved in cellular proliferation (eg.c erbB2 receptor and transcription factor DP2) and a down regulation of genes involved in cell apoptosis (caspase9) or DNA excision repair (ERCC5). While overexpre ssion of HCAP1N resulted in an up regulation of genes involved in cellular suppressors (eg.snoN and WAF1) or DNA excision repair (ERCC5) and a down regulation of genes involved in anti apoptosis (HSPs). Conclusion: Over expression of HCAP1M may promote cell proliferation. Over expression of HCAP1N may suppress cell growth.

关 键 词：HCAP1 基因型 肝癌细胞系 Hep3B基因 表达 

分 类 号：R735.7[医药卫生—肿瘤]