miR-144-3p靶向调控跨膜蛋白16A基因抑制骨肉瘤143B细胞的增殖和侵袭实验

摘要

Objective To identify the expressions of miR-144-3p and transmembrane protein 16 (TMEM16A) in osteosarcoma 143B cells and explore the effect of miR-144-3p which may regulate TMEM16A expression on cell proliferation and invasion.Methods MiR-144-3p was predicted by bioinformatics which may regulate TMEM16A expression. After transfection of miR-144-3p mimics or siRNA against TMEM16A into cells, expressions of miR-144-3p and TMEM16A were detected by real-time PCR or western blot. The proliferation and invasion of 143B cells were respectively determined using MTT assay and transwell chamber assay. Results MiRanda and TargetScan showed that there was a good complementary relationship between miR-144-3p and TMEM16A gene. Results of real-time PCR and western blot showed over-expression of miR-144-3p downregulated the expression levels of TMEM16A mRNA and protein. The proliferation and invasion of 143B cells were suppressed after transfection of miR-144-3p mimics or siRNA against TMEM16A. Conclusion MiR-144-3p may inhibit TMEM16A expression in osteosarcoma 143B cells and subsequently suppress cell proliferation and invasion.%目的 阐明骨肉瘤143B细胞中miR-144-3p与跨膜蛋白16A(transmembrane protein 16,TMEM16A)的表达关系,明确miR-144-3p与143B细胞增殖和侵袭的关系.方法 预测miR-144-3p和TMEM16A基因的互补配对关系,转染miR-144-3p模拟物后,Real-time PCR检测各组癌细胞中miR-144-3p和TMEM16A mRNA的相对表达水平,Western blot检测各组癌细胞中TMEM16A蛋白的相对表达,MTT法检测各组癌细胞的增殖,Transwell检测各组癌细胞体外的侵袭情况.结果 TargetScan和miRanda均显示miR-144-3p与TMEM16A基因互补配对较好.Real-time PCR和Western blot检测结果 表明miR-144-3p的过表达能够降低TMEM16A mRNA和蛋白的相对表达水平.MTT法和Transwell实验发现miR-144-3p的过表达能够抑制143B细胞的增殖和侵袭.结论 miR-144-3p通过下调骨肉瘤143B细胞中TMEM16A的表达水平进而抑制癌细胞的增殖和侵袭.