miR-543调控乳腺癌细胞增殖和侵袭的机制分析

Mechanism Analysis of miR-543 Regulating Breast Cancer Cell Proliferation andInvasion

目的探讨miR-543调控乳腺癌细胞增殖和侵袭的效果与机制。方法对数生长期的人乳腺癌细胞(MDA-MB-468)分为三组:对照组、miR-543组与空白组,对照组、miR-543组分别转染mimics NC与miR-543 mimics,空白组转染等体积的磷酸盐缓冲液。MTT法检测细胞增殖指数,流式细胞仪检测细胞凋亡,Transwell小室检测细胞转移与侵袭,qRT-PCR检测mRNA表达水平,Western Blotting检测蛋白表达水平。结果转染后24 h与36 h,miR-543组的miR-543相对表达水平显著高于空白组与对照组(P<0.05),空白组与对照组对比差异无统计学意义(P>0.05)。转染后24 h与36 h,miR-543组的细胞增殖指数、转移与侵袭指数、B细胞特异性小鼠白血病病毒插入位点1(B-cell-specific Moloney murine leukemia virus insertion site 1,Bmi-1)蛋白相对表达水平显著低于空白组与对照组(P<0.05),细胞凋亡指数显著高于空白组与对照组(P<0.05),空白组与对照组对比差异无统计学意义(P>0.05)。结论miR-543在乳腺癌细胞中可抑制Bmi-1基因的表达,促进细胞凋亡,发挥抑制细胞增殖和侵袭的作用。

Objective To investigate the effect and mechanism of miR-543 on the proliferation and invasion of breast cancer cells.Methods Human breast cancer cells(MDA-MB-468)in logarithmic growth phase were divided into 3 groups:the control group,the miR-543 group and the blank group.The control group and the miR-543 group were transfected with mimics NC and miR-543 mimics,respectively,the blank group was transfected with an equal volume of phosphate buffer.Cell proliferation index were detected by MTT assay,apoptosis were detected by flow cytometry,cell transfer and invasion were detected by Transwell chamber,mRNA expression level were detected by qRT-PCR,and protein expression level were detected by Western Blotting.Results At 24 h and 36 h after transfection,the relative expression level of the miR-543 group were significantly higher than that in the blank group and the control group(P<0.05),and there were no significant difference compared between the blank group and the control group(P>0.05).At 24 h and 36 h after transfection,the relative expression level of B-cell-specific Moloney murine leukemia virus insertion site 1 protein,cell proliferation index,metastasis and invasion index in the miR-543 group were significantly lower than that of the blank group and the control group(P<0.05),and the apoptotic index were significantly higher than that of the blank group and the control group(P<0.05),and there were no significant difference compared between the blank group and the control group(P<0.05).Conclusion miR-543 can inhibit the expression of Bmi-1 gene,promote cell apoptosis and inhibit cell proliferation,and invasion in breast cancer cells.