miR-1284过表达对胃癌细胞中 EIF4 A1的影响

摘要

Objective To explore the potential target genes of miR-1284 and its possible molecular mecha-nisms on gastric cancer development.Methods The bioinformatics software was used to predict the target genes of miR-1284.The slow virus mediated miR-1284 transfection MGC-803 gastric cancer cells were taken as the miR-1284 group(LV-miR-1284).The empty carrier slow virus vector transfection cells were taken as the negative control group and the untransfected slow virus carrier cells as the blank control group.Fluorescence quantitative PCR was used to detect the expression of target genes EIF4A1and Western blot was used to detect EIF4A1 protein expression and its target genes were verified by dual luciferase EIF4A1.Results Compared with those in the negative control group and the blank control group, the expressions of gene and protein of EIF4A1 were decreased in the miR-1284 group, Dual luciferase test demonstrated that miR -1284 could combine with target genes EIF4A1 3′UTR.Conclusion miR-1284 plays a role in biological functions by combining with its target gene EIF4A1.%目的：通过验证miR-1284可能的靶基因，从而探讨miR-1284影响胃癌发生发展可能的分子作用机制。方法采用生物信息学软件预测miR-1284可能的靶基因。以慢病毒介导miR-1284过表达转染胃癌MGC-803细胞为miR-1284组（ LV-miR-1284组），转染空载体慢病毒载体的细胞为阴性对照组，未转染慢病毒载体的细胞为空白对照组。运用荧光定量PCR检测各组EIF4A1基因的表达，Western blot法检测各组EIF4A1蛋白的表达，通过双荧光素酶对EIF4A1进行靶基因验证。结果与阴性对照组和空白对照组比较， miR-1284组的EIF4A1基因和蛋白的表达量下降，双荧光素酶示miR-1284能与靶基因EIF4A1的3′UTR的结合。结论 miR-1284通过作用其靶基因EIF4A1发挥生物功能学作用。