miR-214-3p通过靶向PTEN抑制骨肉瘤U2OS细胞凋亡的研究

miR-214-3p Inhibits Osteosarcoma U2OS Cell Apoptosis by Targeting PTEN

目的:研究骨肉瘤中显著高表达的miR-214-3p对骨肉瘤细胞凋亡的影响及相关作用机制。方法:以骨肉瘤细胞U2OS为研究对象,采用慢病毒感染方法,建立miR-214-3p的稳定转染细胞系,设置过表达negative control慢病毒为对照组,采用流式细胞术检测2组细胞凋亡情况,并对凋亡相关信号通路进行研究。结果:(1)流式结果显示,与对照组比较miRNA-214-3p稳转U2OS细胞系后凋亡细胞减少;(2)Dual-Glo Luciferase方法证实在U2OS细胞中miR-214-3p可以直接结合pten的3'UTR区域,WB实验证实miR-214-3p蛋白水平抑制pten;(3)miR-214-3p可以上调pAKT蛋白表达。结论:miR-214-3p通过直接靶向抑癌基因pten,活化AKT通路,抑制骨肉瘤细胞的凋亡从而在骨肉瘤中发挥了癌基因的作用,可能促进了骨肉瘤的进展。

Objective:To investigate the effect of miR-214-3 p,a highly expressed osteosarcoma,on the apoptosis of osteosarcoma cells and its mechanism.Methods:The osteosarcoma cell line U2OS was used as the research object,and the stable transfected cell line of miR-214-3 p was established by lentivirus method.The apoptosis of U2OS was detected,and the apoptosis related signal pathway was studied.Results:(1)compared with the control,miRNA-214-3 p stable cell apoptosis in U2OS cells decreased;(2)Dual-Glo Luciferase confirmed in U2OS cells miR-214-3 p could directly bind to the 3'UTR region PTEN,WB confirmed that the miR-214-3 p protein level of PTEN is inhibited;(3)miR-214-3 p can increase the expression of pAKT.Conclusion:miR-214-3 p can inhibit the apoptosis of osteosarcoma cells by directly targeting the tumor suppressor gene PTEN and activating AKT pathway,thereby playing the role of oncogene and promoting the progression of osteosarcoma.