摘要
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.
出处
《长治医学院学报》
2017年第6期411-413,共3页
Journal of Changzhi Medical College
