着丝粒蛋白CENP-H对乳腺癌细胞株MCF7增殖能力的影响

目的 研究CENP-H对乳腺癌细胞增殖能力的影响,初步探讨CENP-H与乳腺癌发生、发展的关系.方法 将反转录病毒质粒pMSCV和pMSCV-CENP-H经脂质体转染至293FT细胞制备病毒,并感染MCF7细胞,用嘌呤霉素筛选及Western blot鉴定,建立CENP-H基因稳定表达的MCF7细胞株；应用噻唑盐(MTT)法、平板集落形成实验、5-溴-2-脱氧尿苷(BrdU)掺入法检测CENP-H对MCF7细胞增殖的影响.结果 成功建立稳定表达CENP-H的MCF7细胞株,并发现CENP-H过表达可上调细胞增殖相关分子cyclin D1的表达；MTT、平板克隆实验及Brdu掺入实验结果显示CENP-H过表达后,MCF7的增殖能力模型增强.结论 CENP-H可上调cyclin D1的表达,增强MCF7的增殖能力,提示CENP-H可能在乳腺癌发生、发展中起重要作用.

Effect of CENP-H on proliferation of human breast cancer cell line MCF7

Objective To investigate the effect of CENP-H on proliferation of breast cancer cell line MCF7,and to explore the role of CENP-H in development and progression of breast cancer.Methods Retrovirus vectors pMSCV and pMSCV-CENP-H were transfected into 293FT cells to produce retrovirus.The retrovirus was then infected with the MCF7 cells.The cells were selected with puromycin to establish stable cell lines expressing CENP-H or the control vector.MTT assay,colony formation assay and BrdU incorporation assay were applied to evaluate the proliferation of MCF cells which expressed CENP-H or the control vector.Results Stable cell lines expressing CENP-H or the control vector were successfully established.Overexpression of CENP-H significantly enhanced proliferative activity of MCF7 cells,evidently upregulated the expression of cyclin D1 protein.Conclusion CENP-H can upregulate the expression of cyclin D1,and promote proliferation of breast cancer cell line.These results suggest an important role of CENP-H in the development and progression of breast cancer.