靶向YB-1的microRNA表达载体的构建及其对MB-MDA-231细胞增殖的影响

目的：构建靶向YB-1基因的microRNA表达载体,研究其对乳腺癌细胞MB-MDA231增殖的影响。方法：采用microRNA靶基因预测软件预测可能作用于YB-1基因的micro RNA,构建其过表达载体,利用Lipofectamine2000转染细胞MB-MDA231,荧光显微镜下评估转染效果,real-time ...

MicroRNA137 suppressed human breast cancer cell MB-MDA231 proliferation by directly targeting YB-1 gene

AIM：To investigate the inhibition effects of microRNA targeting Y-box binding protein 1（YB-1）gene on the proliferation of human breast cancer cell MB-MDA231.Methods：A series of bioinformatics analysis were performed to predict possible microRNA targeting the 3 UTR of YB-1 gene.According to the results from bioinformatics analysis,the microRNA overexpression pGensill-pre-mir137 plasmid was constructed.The vector was transfected into MB-MDA231 cells by lipofectamine,and the expression of microRNA was detected by real-time PCR.Then the expressions of YB-1 mRNA and protein were analyzed by real-time PCR and Western blot analysis respectively.To evaluate the relationship of microRNA with 3＇UTR of YB-1 gene,the dual luciferase assays were performed,and MTT assays were used to observe the inhibitory effect of microRNA on cell proliferation in vitro.Results：pGensil 1-pre-mir 137 plasmid was successfully constructed and identified by enzyme restriction digestion and sequence analysis.Compared with control groups,the expression of microRNA137 was increased remarkably（P0.05）, the expression of YB-1 mRNA and protein level were significantly reduced（P 0.05）.In MB-MDA231 cotransfected by pGensill-pre-mir137 and pMIR-Luc-YB1 plasmids,luciferase reporter activities reduced by 30% compared with that in MB-MDA231 cotransfected by pGensill and pMIR-Luc-YB 1 plasmids.The growth ability of cells in experimental group were lower than that of control groups（P0.05）.Conclusions The microRNA137 overexpression plasmid is successfully constructed,and microRNA 137 can suppress human breast cancer cell proliferation by directly targeting 3＇ UTR of YB-1 gene.