【摘要】 目的探讨miR-148a-3p在乳腺癌中的作用及可能的机制。方法根据人乳腺癌细胞株-7(MCF-7)细胞是否转染及是否存在miR-148a-3p的模拟物分为3个组:未转染组、miR-148a-3p对照组和miR-148a-3p转染组。通过实时荧光定量PCR(RT-qPCR)检测转染后miR-148a-3p在各组的信使RNA(mRNA)水平,运用四唑盐比色法(MTT)比较MCF-7细胞在3组的增殖情况,应用RT-qPCR检测各组神经母细胞瘤病毒性ras致癌因子的同原体(NRAS)的基因水平,应用Western blot检测各组RAS病毒致癌基因同源物NRAS的蛋白水平,应用流式细胞术检测各组细胞凋亡情况,应用双荧光素酶报告基因实验验证miR-148a-3p与NRAS之间的作用。结果过表达miR-148a-3p显著抑制乳腺癌细胞株MCF-7的增殖(P<0.05),促进其凋亡(P<0.05),降低NRAS基因和蛋白的表达(P<0.05),双荧光素酶报告基因实验证明NRAS是miR-148a-3p的直接作用靶点。结论 miR-148a-3p可能通过靶向NRAS抑制乳腺癌MCF-7细胞的增殖,促进细胞凋亡。更多还原

【Abstract】 Objective To investigate the role of miR-148 a-3 p in breast cancer and its possible molecular mechanism.Methods According to whether human breast cancer cell line-7(MCF-7)cells were transfected and whether miR-148 a-3 p mimics were present,MCF-7 cells were divided into three groups:the untransfected group,the control group and the transfected group.The real-time fluorescent quantitative PCR(RT-qPCR)was used to assess the messenger RNA(mRNA)level,the proliferation of MCF-7 cells were conducted by methyl thiazolyl tetrazolium(MTT),RT-qPCR was used to detect the gene level of ras virus oncogene homolog(NRAS),the protein level of NRAS was detected by Western blot,the apoptosis was detected by cytometry,and the effect of miR-148 a-3 p and NRAS was verified by dual luciferase reporter gene assay.ResultsOverexpression of miR-148 a-3 p significantly inhibited the proliferation of MCF-7 cells(P<0.05),promoted the apoptosis of the cells(P<0.05),decreased the expression of NRAS gene and protein(P<0.05),dual luciferase reporter gene experiments demonstrated that NRAS was a direct target of miR-148 a-3 p.ConclusionmiR-148 a-3 p inhibits the proliferation and induces the apoptosis of MCF-7 cells by inhibiting the expression of NRAS.更多还原