miR-25-3p调控CPEB4基因对人肺癌A549细胞侵袭和迁移的影响

Effect of miR-25-3p on invasion and migration of human lung cancer A549 cells by regulating expression of cytoplasmic polyadenylation element binding protein 4

目的探讨miR-25-3p和CPEB4基因在人肺癌A549细胞中的表达,阐明miR-25-3p对CPEB4基因的靶向作用及其对A549细胞侵袭和迁移的影响。方法采用免疫荧光化学技术检测CPEB4在A549细胞中的表达;运用生物信息学方法对miR-25-3p和CPEB4基因的靶向配对关系进行预测;脂质体2000转染miR-25-3p模拟物以及干扰CPEB4基因的siRNA后,采用Real-time PCR检测miR-25-3p和CPEB4基因mRNA在癌细胞中的表达;Western blot法检测CPEB4蛋白在癌细胞中的表达;Transwell小室试验检测癌细胞体外的侵袭性;细胞划痕试验检测癌细胞的迁移情况。结果 A549细胞胞质内CPEB4蛋白高表达,呈绿色荧光;miR-25-3p和CPEB4基因二者靶向配对良好;过表达miR-25-3p能降低A549细胞中CPEB4基因mRNA和蛋白的表达,抑制A549细胞的侵袭和迁移。结论 miR-25-3p通过下调人肺癌A549细胞中CPEB4基因的表达,进而抑制癌细胞的侵袭和迁移。

Objective To investigate the expression of miR-25-3p and cytoplasmic polyadenylation element binding protein 4（CPEB4） in human lung cancer A549 cells, the targeting effect of miR-25-3p to CPEB4 gene as well as the effect of miR-25-3p on invasion and migration of A549 cells. Methods The expression of CPEB4 in A549 cells was determined by immunofluorescence cytochemical assay. The targeting effect of MiR-25-3p to CPEB4 gene was predicted by bioinformatics. The expressions of miR-25-3p and CPEB4 mRNAs in A549 cells transfected with miR-25-3p mimics and siRNA in mediation of liposome 2000 were determined by real-time PCR, while that of CPEB4 by Western blot. The invasion of A549 cells was determined by Transwell chamber test, while the migration by wound healing assay. Results CPEB4 was highly expressed in the cytoplasm of A549 cells, which showed green fluorescence. The miR-25-3p was well complementary with CPEB4 gene. The overexpression of miR-25-3p down-regulated the expressions of CPEB4 mRNA and protein, and inhibited the invasion and migration of A549 cells. Conclusion The miR-25-3p may down-regulate the CPEB4 expression in human lung cancer A549 cells and inhibit the cell invasion and migration.