微小RNA-382-5p(miR-382-5p)对乳腺癌细胞增殖和凋亡的影响及作用机制

Effects of microRNA-382-5p(microRNA-382-5p)on Proliferation and Apoptosis of Breast Cancer Cells and Its Mechanism

目的观察微小RNA-382-5p(miR-382-5p)对乳腺癌细胞增殖和凋亡的影响并探讨其作用机制。方法选择乳腺癌患者86例,取患者乳腺癌组织0.5 cm×0.5 cm×0.5 cm为观察组,同时取与乳腺癌病灶间隔5 cm以上同样大小的正常组织为对照组。行细胞培养和转染、PT-PRC检测、Western blot法检测蛋白表达、平板克隆试验、CCK-8法检测和流式细胞术。结果观察组样本miR-382-5p与对照组相比呈现显著低表达,且具有统计学差异(P<0.05),观察组转染后的MCR-7细胞miR-382-5p表达量高于对照组,且具有统计学差异(P<0.05),但观察组细胞中NF90 mRNA较对照组明显降低,且均具有统计学差异(P<0.05),细胞转染48 h后,Western blot法检测结果显示MCF-7细胞转染miR-382-5p后NF90、Bcl-2蛋白表达观察组较对照组明显下调,p53、p21、Bax观察组较对照组明显上调,Cyclin E1蛋白无明显变化。观察组患者克隆形成数低于对照组,细胞凋亡率高于对照组,且具有统计学差异(P<0.05),采用CCK法检测不同时间点MCF-7细胞生长,在细胞生长第4、5天,观察组细胞活力低于对照组,且具有统计学差异(P<0.05),细胞生长1、2、3天两组细胞活力无统计学差异(P<0.05),经过流式细胞术检查,显示MCF-7细胞转染miR-382-5p后凋亡率高于对照组,差异具有统计学意义(P<0.05)。结论miR-382-5p具有抑制乳腺癌细胞增殖,促进细胞凋亡的作用,其基因与靶向抑制NF90基因具有相关性。

Objective To observe the effect of microRNA-382-5p(microRNA-382-5p)on proliferation and apoptosis of breast cancer cells and explore its mechanism.Methods 86 patients with breast cancer diagnosed were selected.The breast cancer tissues of 0.5 cm×0.5 cm×0.5 cm were taken as the observation group,and the normal tissues of the same size as breast cancer lesions with intervals of more than 5 cm were taken as the control group.Cell culture and transfection,PT-PRC detection,Western blot detection of protein expression,plate cloning test,CCK-8 detection and flow cytometry were performed.Results The expression of microRNA-382-5p in MCR-7 cells in the observation group was significantly lower than that of the control group(P<0.05).The expression of microRNA-382-5p in MCR-7 cells in the observation group was higher than that of the control group(P<0.05),but the expression of NF90 in the observation group was significantly lower than that of the control group(P<0.05).The results of RN blot showed that the expression of NF90 and Bcl-2 protein in MCF-7 cells transfected with microRNA-382-5p was significantly lower in the observation group than in the control group,while the expression of p53,p21 and Bax in the observation group was significantly higher than that in the control group,while the expression of Cyclin E1 protein had no significant change.The cell viability of MCF-7 cells in the observation group was lower than that in the control group on the 4th and 5th day of cell growth,and there was no significant difference in cell viability between the 2 groups on the 1st,2nd and 3rd day of cell growth(P>0.05).Flow cytometry showed that the apoptotic rate of MCF-7 cells transfected with microRNA-382-5p was higher than that of the control group(P<0.05).Conclusion Mi-382-5p can inhibit the proliferation of breast cancer cells and promote cell apoptosis,and its gene is related to the targeting inhibition of NF90 gene.