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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。
Objective To investigate the effect of CENP-H on the proliferation of breast cancer cells and to explore the relationship between CENP-H and the occurrence and development of breast cancer. Methods The retroviral plasmids pMSCV and pMSCV-CENP-H were transfected into 293FT cells by lipofectamine to prepare virus and MCF7 cells were infected by puromycin and identified by Western blot. The CENF-H gene was stably expressed in MCF7 Cell lines. The effects of CENP-H on the proliferation of MCF7 cells were detected by MTT assay, platelet colony formation assay and BrdU incorporation assay. Results CENP-H stably expressing MCF7 cell line was successfully established and found that CENP-H overexpression up-regulated cyclin D1 expression. MTT assay, plate clone assay and Brdu incorporation assay showed that CENP-H over- MCF7 proliferation model enhanced. Conclusion CENP-H up-regulates the expression of cyclin D1 and enhances the proliferation of MCF7, suggesting that CENP-H may play an important role in the occurrence and development of breast cancer.