摘要
Abstract
Objective To investigate the effect of miR-130a-5p targeting tumor protein D52(TPD52)on proliferation and apoptosis of human breast cancer cells.Methods Human breast cancer MCF-7 cells were randomly divided into the control group,the NC mimic group(transfected with mimic negative control),the miR-130a-5p mimic group(transfected with miR-130a-5p mimic),the miR-130a-5p mimic+pcDNA-NC group(co-transfected with miR-130a-5p mimic and pcDNA negative control),and the miR-130a-5p mimic+pcDNA-TPD52 group(co-transfected with miR-130a-5p mimic and pcDNA-TPD52).Bioinformatics software and dual-luciferase reporter assay were used to analyze and validate the targeting relationship of miR-130a-5p to TPD52.Quantitative real-time PCR was used to detect the mRNA expression of miR-130a-5p and TPD52 mR-NA in each group.Cell counting kit-8(CCK-8)assay was used to assess cell proliferation.Flow cytometry and 4',6-diamidino-2-phenylindole(DAPI)nuclear staining were used to detect apoptosis rate of cells and nuclear apoptosis.Western blot was used to detect the expression of proliferation-related and apoptosis-related pro-teins.Results There were consecutive binding sites between miR-130a-5p and the 3'-UTR of TPD52,and miR-130a-5p could target and regulate TPD52 expression.Compared with the NC mimic group,the relative expression level of miR-130a-5p in the miR-130a-5p mimic group was significantly increased,while the relative expression level of TPD52 mRNA was significantly decreased(P<0.05).Compared with the miR-130a-5p mimic+pcDNA-NC group,the relative expression level of TPD52 mRNA in the miR-130a-5p mimic+pcDNA-TPD52 group was significantly increased(P<0.05).Compared with the NC mimic group,the cell viability in the miR-130a-5p mimic group was significantly decreased(P<0.05),while the apoptosis rate was significant-ly increased(P<0.05).Compared with the miR-130a-5p mimic+pcDNA-NC group,the cell viability in the miR-130a-5p mimic+pcDNA-TPD52 group was significantly increased(P<0.05),while the apoptosis rate was significantly decreased(P<0.05).Compared with the NC mimic group,the expression levels of Bax and caspase 3 proteins in the miR-130a-5p mimic group were significantly increased(P<0.05),while the expres-sion level of PCNA protein was significantly decreased(P<0.05).Compared with the miR-130a-5p mimic+pcDNA-NC group,the expression levels of Bax and caspase 3 proteins in the miR-130a-5p mimic+pcDNA-TPD52 group were significantly decreased(P<0.05),while the expression level of PCNA protein was signifi-cantly increased(P<0.05).Conclusion miR-130a-5p inhibits MCF-7 cell proliferation and induces apoptosis by targeting TPD52.关键词
miR-130a-5p/肿瘤蛋白D52/乳腺癌/增殖/凋亡Key words
miR-130a-5p/tumor protein D52/breast cancer/proliferation/apoptosis分类
医药卫生
