四环素诱导表达CARM1在乳腺癌细胞中的生物学效应

The Effects of the Tetracyclin-induced Expression of CARM1 in Breast Cancer Cells

为了探究共激活因子相关的精氨酸甲基转移酶1(CARM1)在乳腺癌细胞中的生物学作用,构建了四环素诱导过表达CARM1的乳腺癌MDA-MB-231细胞株,并应用蛋白质印迹法(Western blot)、实时荧光定量聚合酶链式反应(RT-PCR)对相关指标进行检测。结果发现,在乳腺癌细胞株中成功诱导CARM1过表达之后,CARM1蛋白水平和mRNA水平均显著升高。此外,利用细胞划痕、平板克隆、细胞增殖试验、流式细胞术等试验方法检测过表达CARM1对细胞迁移、增殖、克隆形成以及细胞周期进程的影响。试验结果显示,诱导CARM1过表达后,细胞迁移能力和克隆形成能力均明显增强,同时细胞增殖试验表明细胞的增殖能力也明显增强。流式细胞术分析发现,诱导表达CARM1后,细胞株S期和G2/M期的细胞比率增加,过表达CARM1能够促进细胞周期的进程。以上研究结果提示,在乳腺癌细胞中过表达CARM1可以促进细胞的增殖、提高细胞的克隆形成及迁移的能力,加快细胞周期进程。该结论为后续CARM1在乳腺癌细胞中的相关研究奠定了基础。

In order to investigate the biological roles of CARM1 in breast cancer,we constructed MDA-MB-231 breast cancer cell line overexpressing CARM1 via tetracycline induction in this study.Western blot and RT-PCR were employed to determine the significant parameters.We revealed that the levels of both CARM1 protein and mRNA significantly increased after successful induction of CARM1 expression.In addition,the effects of overexpression of CARM1 on cell migration,proliferation,clonal formation and cell cycle were analyzed by cell scratch assay,plate cloning assay,cell proliferation assay,and flow cytometry.The results exhibited that the overexpression of CARM1 enhanced cell migration ability,clonal formation ability and cell proliferation ability of MDA-MB-231 cells.Moreover,the proportion of cells at S and G2/M phases were elevated in CARM1-overexpressed samples detected by flow cytometry analysis,implicating that the overexpression of CARM1 promoted the cell cycle process.In conclusion,this study suggested that the overexpression of CARM1 could promote the proliferation of cells,improve the ability of colony formation and migration,and accelerate the cell cycle process in MDA-MB-231 cells,which can serve as a foundation for further studies on CARM1 in breast cancer cells.