miRNA-183-5p在乳腺癌患者的表达及对乳腺细胞上皮间质转化、迁移和侵袭的影响

Expression of miRNA-183-5p in patients with breast cancer and its effects on breast cancer cell epithelial-mesenchymal transition,migration and invasion

目的研究miRNA-183-5p在乳腺癌患者体内的表达情况及对乳腺细胞上皮间质转化的影响。方法采用Real-timePCR检测93例乳腺癌患者的乳腺癌组织和癌旁组织以及乳腺癌细胞MDA-MB-231和正常乳腺细胞MCF-10A中miRNA-183-5p的表达水平。将MDA-MB-231分为3组:未做处理的对照组,转染control miRNA的mimics control组以及转染miRNA-141-5p mimics的mimics组,Real-time PCR法检测3组细胞中miRNA-183-5p的表达水平;采用Western blot方法检测基质金属蛋白酶-9(MMP9),上皮细胞-间质细胞转化(EMT)相关的E-钙黏蛋白(E-cadherin)及波形蛋白(Vimentin)的表达情况;划痕愈合实验检测细胞迁移能力;Transwell小室法检测细胞侵袭能力。结果miRNA-183-5p在乳腺癌组织、MDA-MB-231细胞中的表达水平显著低于癌旁组织及MCF-10A细胞(P<0.05)。mimics组的细胞中miRNA-183-5p的表达水平显著高于对照组和mimics control组(P<0.05)。E-cadherin在mimics组的水平显著高于对照组和mimicscontrol组(P<0.05),MMP-9及Vimentin在mimics组的水平显著低于对照组和mimicscontrol组(P<0.05)。与其他2组比较,mimics组的划痕愈合率以及侵袭能力显著降低(P<0.05)。结论miRNA-183-5p在乳腺癌患者体内呈低水平表达,可能通过靶向降低MMP-9的表达水平,抑制乳腺癌细胞MDA-MB-231的上皮间质转化、迁移和侵袭能力。

Objective To investigate the expression of miRNA-183-5p in patients with breast cancer and its effects on breast cancer cell epithelial-mesenchymal transition,migration and invasion.`Methods The Real-time PCR was used to detect the expression levels of miRNA-183-5p in breast cancer tissues and the corresponding normal tissues from 93 patients with breast cancer,as well as the expession levels were detected in breast cancer cell-MDA-MB-231 and normal mammary cell-MCF-10 by Real-time PCR.MDA-MB-231 cells were divided into 3 groups:the cells without any treatment were enrolled as control group,and the cells were transfected with control miRNA were enrolled as mimicscontrol group,and the cells were transfected with miRNA-183-5p mimics were enrolled as mimics group.The expression levels of miRNA-141-5p in the three groups were detected by Real-time PCR.Moreover the expression levels of E-cadherin and Vimentin were detected by Western Blot to verify the EMT,and the cell migration was tested by scratch healing experiment,and the cell invasion was tested by Transwell experiment.In addition the expression levels of matrix metalloproteinase-9(MMP-9)were detected.Results The expression levels of miRNA-183-5p in breast cancer tissues and MDA-MB-231 cells were significantly lower than those in corresponding normal tissues and MCF-10A cells(P<0.05).The expression levels of miRNA-183-5p in mimics group were significantly higher than those in mimicscontrol group and control group(P<0.05).The expression levels of MMP-9 and E-cadherin in mimicsgroup were significantly higher than those in mimicscontrol group and control group(P<0.05).Scratch healing rate and cell invasion ability in mimics group were significantly lower than those in the other two groups(P<0.05).Conclusion The miRNA-183-5p is poorly expressed in patients with breast cancer,which may inhibit EMT,migration and invasion of MDA-MB-231 cells by targetedly reducing the expression levels of MMP-9.