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目的:探讨MST1(mammalian sterile 20-like kinase 1)基因对人乳腺癌细胞MCF-7增殖与凋亡的影响。方法:构建含有标签蛋白FLAG的MST1质粒pCMV-FLAG-MST1;将构建好的pCMV-FLAG-MST1质粒在脂质体lipofectamine 2000的介导下转染MCF-7细胞,通过Western印迹法分析MST1在MCF-7中的表达情况;分别在转染后12、24、36和48h通过MTT法观察MST1对细胞增殖的影响;转染后36h加入5-溴-2’脱氧尿嘧啶(bromodeoxy uridine,BrdU),通过其掺入比率显示MST1对细胞增殖的影响;转染后36h加入顺铂,作用14h后通过AnnexinⅤ检测其对细胞凋亡的影响。结果:成功构建pCMV-FLAG-MST1质粒;MTT及BrdU检测显示,转染pCMV-FLAG-MST1质粒后MCF-7细胞增殖明显受到抑制;Annexin Ⅴ检测结果提示,高表达MST1后,细胞的凋亡率相对增高。结论:在乳腺癌细胞MCF-7中高表达MST1,不仅可以抑制细胞增殖,还可以促进细胞凋亡。
Objective: To investigate the effect of MST1 gene on the proliferation and apoptosis of human breast cancer cell line MCF-7. Methods: The MST1 plasmid pCMV-FLAG-MST1 containing the tagged protein FLAG was constructed. The constructed pCMV-FLAG-MST1 plasmid was transfected into MCF-7 cells by lipofectamine 2000 and analyzed by Western blotting MCF-7 in transfected cells were detected by MTT assay. The effect of MST1 on cell proliferation was observed at 12,24,36 and 48 h after transfection. BrdU (BrdU) ). The effect of MST1 on cell proliferation was shown by its incorporation rate. Cisplatin was added 36h after transfection and the effect of MST1 on apoptosis was detected by Annexin V after 14h. Results: The pCMV-FLAG-MST1 plasmid was constructed successfully. The MTT assay and BrdU assay showed that the proliferation of MCF-7 cells was obviously inhibited after transfected with pCMV-FLAG-MST1 plasmid. Annexin Ⅴ test showed that MST1- The rate is relatively high. Conclusion: High expression of MST1 in breast cancer cell line MCF-7 not only can inhibit cell proliferation, but also promote apoptosis.