miR-590-5 p靶向调控STAT3基因对骨肉瘤143 B细胞迁移和侵袭的影响

摘要

目的:探讨miR-590-5p和信号转导与转录激活因子3(STAT3)基因在骨肉瘤143B细胞中的表达,探讨miR-590-5p对STAT3基因的靶向作用及其对143B细胞迁移和侵袭的影响.方法:运用生物信息学方法对miR-590-5p和STAT3基因的靶向配对关系进行预测,采用荧光素酶报告系统鉴定;脂质体2000转染miR-590-5p模拟物以及干扰STAT3的siRNA后,real-time PCR检测miR-590-5p和STAT3 mRNA在癌细胞中的表达,Western blotting检测STAT3蛋白在癌细胞中的表达,细胞划痕实验检测癌细胞的迁移情况以及Transwell小室检测癌细胞体外的侵袭性.结果:生物信息学软件TargetScan和miRanda显示miR-590-5p和STAT3基因二者靶向配对良好,荧光素酶报告系统鉴定发现miR-590-5p能够抑制STAT3 mRNA表达.Real-time PCR和Western blotting检测结果均表明过表达miR-590-5p能够降低STAT3 mRNA和蛋白的表达.细胞划痕实验和Transwell实验发现过表达miR-590-5p能够分别抑制143B细胞的迁移和侵袭.结论:miR-590-5p通过负性靶向调控骨肉瘤143B细胞中STAT3基因的表达,进而抑制癌细胞的迁移和侵袭.%Objective:To explore the expressions of miR-590-5p and STAT3 in osteosarcoma 143B cells,and the effects of STAT3 expression regulated by miR-590-5p on the migration and invasion of osteosarcoma 143B cells. Methods:The target pairing of miR-590-5p and STAT3 genes was predicted by bioinformatics, and identified with luciferase assay. After miR-590-5p mimics and siRNA targeting STAT3 transfecting into cells using lipofectamine 2000,the expressions of miR-590-5p and STAT3 gene were determined by real-time PCR,and the STAT3 protein was determined using Western blotting. The migration and invasion in vitro of cancer cells were detected using the wound healing assay and transwell chamber,respectively. Results:The results of MiRanda and TargetScan showed that the target pairing of miR-590-5p and STAT3 genes was good. The results of luciferase assay showed that MiR-590-5p could inhibit the expression of STAT3 mRNA. The results of real-time PCR and Western blotting showed the over-expression of miR-590-5p could down-regulated the expressions of STAT3 mRNA and protein. The migration and invasion of 143B cells were suppressed after transfecting miR-590-5p mimics. Conclusions:MiR-590-5p can inhibit the migration and invasion of osteosarcoma 143B cells by negative regulating the STAT3 expression.