摘要
目的探讨ADAM17在U87MG细胞增殖与侵袭中的作用及机制。方法收集人脑胶质瘤组织65例(肿瘤组)与正常脑组织13例(对照组),采用免疫组化、RT-PCR法,检测ADAM17蛋白及mRNA的表达;利用siRNA干扰敲减或ADAM17激活剂(PMA)过表达ADAM17,并予以PI3K抑制剂抑制相关信号通路,MTT及Transwell实验检测各组U87MG细胞的增殖与侵袭变化,Western blot检测ADAM17、p-AKT及AKT蛋白变化。结果胶质瘤组织中ADAM17mRNA及蛋白水平均高于正常脑组织;与对照组相比,siRNA干扰后,低表达组U87MG细胞的增殖与侵袭能力下降,而激活剂PMA增强ADAM17后,过表达组细胞增殖与侵袭能力增加,差异有统计学意义(P<0.05);敲减及过表达ADAM17可导致PI3K/AKT信号通路下游蛋白发生变化。结果 ADAM17可通过激活PI3K/AKT信号通路促进U87MG细胞增殖与侵袭,有望为胶质瘤的治疗找到新突破点。
Objective To investigate role and mechanism of ADAM17-mediated proliferation and invasion of U87 MG glioma cell. Methods The expression levels of ADAM17 in 65 cases of gliomas and 13 cases of normal brain tissues were detected by immunohistochemical staining and RT-PCR. The proliferation and invasion ability of U87 MG cells were detected by MTT and transwell assays after silencing ADAM17 by siRNA or overexpression ADAM17 with the ADAM17 agonist（ PMA）. Western Blot was used to detect ADAM17,AKT and p-AKT protein expression of U87 MG cells in each group. Results ADAM17 mRNA and protein expressions were both higher in glioma than that in normal brain tissues（ P〈0. 05）; Compared with the control group,stimulation of U87 MG cells with the ADAM17 agonist（ PMA） improved the proliferation and invasion ability,while knockdown of ADAM17 in U87 MG cells by si-ADAM17 decreased the proliferation and invasion ability（ P〈0. 05）. Knock-down or overexpression ADAM17 can lead to p-AKT protein changes of PI3K/AKT signaling pathway. Conclusion ADAM17 could mediate the proliferation and invasive of U87 MG glioma cell by activating PI3K/AKT signaling pathway. It is expected to find a potential therapeutic target for the treatment of glioma.
出处
《实用癌症杂志》
2017年第6期883-887,共5页
The Practical Journal of Cancer
基金
南充市科学技术和知识产权局项目(编号:14A0040)
