着丝粒蛋白-H对人胃癌细胞增殖的影响  

Role of centromere protein H in human gastric cancer cell proliferation

作　　者：全天一[1] 何本夫[2] 刘铁坚 李伟明[1] 伍尚标[4] 江庆萍[5] 刘维强 刘海波 徐学虎[4] 

机构地区：[1]广州医学院第三附属医院肿瘤科,广东广州510150 [2]解放军第四二一医院肿瘤科,广东广州510318 [3]广州朗日生物技术有限公司,广东广州510663 [4]广州医学院第三附属医院普通外科,广东广州510150 [5]广州医学院第三附属医院病理科,广东广州510150 [6]广州市生殖与遗传研究实验室,广东广州510150

出　　处：《南方医科大学学报》2012年第2期265-269,共5页Journal of Southern Medical University

基　　金：广州市科技和信息化局科技支撑计划项目(2010J-E141);广东省卫生厅(A2011277)血液中miRNA用于结直肠癌早期诊断的前期研究

摘　　要：目的检测着丝粒蛋白-H(CENP-H)基因在人胃癌细胞系中的表达情况,研究CENP-H对胃癌细胞增殖的影响。方法采用QPCR、Western blot检测7株胃癌细胞系及永生化人胃粘膜上皮细胞中CENP-H的mRNA和蛋白表达水平。用重组逆转录病毒感染胃癌细胞,建立稳定干扰内源性及高表达外源性CENP-H的胃癌细胞系,并利用Western blot及QPCR检测进行鉴定,最后采用MTT、平板克隆形成实验分析CENP-H对胃癌细胞增殖能力的影响。结果人胃癌细胞系中CENP-H蛋白及mRNA表达水平均高于永生化人胃粘膜上皮细胞。Western blot及QPCR鉴定结果表明成功建立稳定沉默内源性及高表达外源性CENP-H的胃癌细胞系。MTT、平板克隆形成实验显示干扰CENP-H后对胃癌细胞增殖能力起到抑制作用,过表达CENP-H后能促进胃癌细胞增殖。结论 CENP-H对胃癌细胞的增殖具有重要的作用,在胃癌的发生发展中可能扮演重要角色。Objective To explore the role of centromere protein H（CENP-H） in the proliferation of human gastric cancer cells. Methods RT-PCR and Western blot analysis were employed to examine the mRNA and protein expressions of CENP-H in 7 human gastric cancer cell lines and immortalized human gastric epithelial cells（GES-1）.The cells were infected with the retrovirus vectors pMSCV-CENP-H or CENP-H-RNAi to establish stable cell lines with high CENP-H expression or CENP-H expression interference.MTT assay and colony formation assay were used to examine the changes in the cell proliferation after the infection.Results CENP-H was over-expressed in gastric cancer cell lines AGS,BGC823,SGC-7901,MKN45,HGC27, MGC-803 and MKN28 at both mRNA and protein levels.The established AGS/CENP-H cell line with increased CENP-H expression showed enhanced proliferative activity,while the cell line MGC-803/CENP-H-RNAi with CENP-H expression interference showed an obviously lowered proliferation ability.Conclusion CENP-H promotes the proliferation of human gastric cancer cells,suggesting its important role in the occurrence and development of gastric cancer.

关 键 词：着丝粒蛋白-H 胃癌 增殖 逆转录病毒 稳定细胞系 

分 类 号：R735.2[医药卫生—肿瘤]