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

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.