miR-328-3p-Akt/mTOR轴在雷公藤甲素抑制脑胶质瘤细胞生长中的作用及机制研究

Role and mechanism of miR-328-3p-Akt/mTOR axis in the inhibition of glioma cell growth by triptolide

目的 以U251细胞和原位胶质瘤小鼠模型为研究对象,观察miR-328-3p对脑胶质瘤细胞生长和凋亡的影响,探讨miR-328-3p-Akt/mTOR轴在雷公藤甲素抑制脑胶质瘤细胞生长中的作用及机制。方法 在U251细胞中过表达miR-328-3p,再结合雷公藤甲素的处理,采用qRT-PCR、CCK8、克隆形成实验、流式细胞术、Western Blot等方法检测miR-328-3p在雷公藤甲素抑制脑胶质瘤细胞U251生长中的作用;构建miR-328-3p稳定过表达的原位胶质瘤小鼠模型,采用qRT-PCR和小动物活体成像体内验证miR-328-3p在雷公藤甲素抑制脑胶质瘤细胞生长的作用。结果 miR-328-3p在脑胶质瘤细胞和组织中表达降低,其表达水平与原发性和复发性脑胶质瘤患者总生存率呈正相关;miR-328-3p与雷公藤甲素可以协同作用抑制脑胶质瘤细胞的生长,促进脑胶质瘤细胞的凋亡,抑制Akt/mTOR信号通路的激活。结论 miR-328-3p在脑胶质瘤细胞中起到抑癌基因的作用,雷公藤甲素可能通过miR-328-3p促进脑胶质瘤细胞凋亡,抑制Akt/mTOR信号通路的激活,进而抑制脑胶质瘤细胞的生长。

Objective To observe the effect of miR-328-3p on the growth and apoptosis of glioma cells,and explore the role and mechanism of the miR-328-3p-Akt/mTOR axis in inhibition of glioma cell growth by triptolide using U251 cells and a xenograft model in nude mice.Methods miR-328-3p overexpression were induced after treatment of with triptolide in U251 cells.qRT-PCR,CCK-8,colony formation assays,flow cytometry,and Western Blot were performed to assess the role of miR-328-3p in inhibition of U251 cells by triptolide.A xenotransplanted tumor model was established with stable miR-328-3p expression to verify the role of miR-328-3p in inhibition of glioma cells by triptolide using qRT-PCR and small animal imaging in vivo.Results miR-328-3p expression in glioma cells and tissues was decreased,and its expression level was positively correlated to the overall survival rate of patients with primary and recurrent glioma.miR-328-3p cooperated with triptolide to inhibit glioma cell growth,promote glioma cell apoptosis,and inhibit activation of the Akt/mTOR signaling pathway.Conclusions miR-328-3p is a tumor suppressor gene in glioma.Triptolide promotes apoptosis of glioma cells,inhibits activation of the Akt/mTOR signal pathway,and inhibits glioma cell growth through miR-328-3p.