雷帕霉素通过活化MAPK促进神经胶质瘤U87-MG细胞自噬  

Rapamycin promotes autophagy of glioma U87-MG cells by activatingMAPK

作　　者：曹相玫 徐辉 武艳玮 游南林 于佳翔 马杰 CAO Xiang-mei;XU Hui;WU Yan-wei;YOU Nan-lin;YU Jia-xiang;MA Jie(Department of Pathology,Basic Medical College,Ningxia Medical University,Ningxia 750004,China)

机构地区：[1]宁夏医科大学基础医学院病理学系,宁夏银川750004

出　　处：《中国病理生理杂志》2021年第4期654-660,共7页Chinese Journal of Pathophysiology

基　　金：宁夏自然科学基金(No.2020AAC03185)。

摘　　要：目的:探讨雷帕霉素(Rapa)对神经胶质瘤U87-MG细胞自噬的相关作用和机制。方法:用Rapa处理U87-MG细胞,分为正常对照(NC)组(0μmol/L Rapa)及Rapa各浓度组(1、2.5、5、10、20和40μmol/L Rapa),MTT实验检测细胞活性,软琼脂集落形成实验和平板集落形成实验检测细胞集落形成能力,Western blot法检测mTOR、p-mTOR、LC3-Ⅱ、beclin-1、p-S6、p70S6K、p-MAPK及GFAP蛋白水平的变化。结果:MTT实验结果显示,Rapa显著抑制U87-MG细胞的活力,并呈现剂量依赖性(P<0.05)。软琼脂和平板集落形成实验结果显示,Rapa显著抑制U87-MG细胞集落形成数(P<0.05)。Western blot结果显示,Rapa处理后mTOR及p-mTOR蛋白水平降低;自噬标志蛋白LC3-Ⅱ和beclin-1水平显著升高(P<0.05);自噬相关信号通路分子p70S6K表达趋势呈抛物线轨迹,差异无统计学意义(P>0.05);p-S6表达趋势与p70S6K相反,Rapa干预后p-S6表达显著降低,在40μmol/L Rapa干预后上升至NC组的水平;p-MAPK和GFAP水平显著增高(P<0.05)。结论:Rapa通过激活MAPK促进细胞自噬,从而抑制U87-MG细胞增殖。AIM:To investigate the effect of rapamycin(Rapa)on autophagy of glioma U87-MG cells.METHODS:Glioma U87-MG cells were treated with Rapa,and were divided into normal control(NC)group(0 μmol/L Rapa)and experimental groups(1,2. 5,5,10,20 and 40 μmol/L Rapa). MTT assay was used to measured the cell viability,and soft agar colony formation assay and plate colony formation assay were used to detect the colony-forming ability of the cells. The protein levels of mTOR,p-mTOR,LC3-Ⅱ,beclin-1,p-S6,p70 S6 K,p-MAPK and GFAP were determined by Western blot.RESULTS:The results of MTT assay showed that Rapa significantly inhibited the viability of U87-MG cells in a dose-dependent manner(P<0. 05). The results of soft agar and plate colony formation assays showed that Rapa significantly inhibited the colony formation of U87-MG cells(P<0. 05). After treatment with Rapa,the protein levels of mTOR and p-mTOR were decreased,the autophagy marker proteins LC3-Ⅱ and beclin-1 were significantly increased(P<0. 05),and the expression of autophagy-related signaling pathway molecules p70 S6 K showed a parabola-like trajectory without statistical significance(P>0. 05). The expression trend of p-S6,opposite to that of p70 S6 K,was decreased significantly after Rapa intervention and increased to the level of NC group after 40 μmol/L Rapa treatment. The protein levels of p-MAPK and GFAP were increased after treatment with Rapa(P<0. 05).CONCLUSION:Rapamycin inhibits the proliferation of U87-MG cells by activating MAPK to promote autophagy.

关 键 词：神经胶质瘤 自噬 雷帕霉素 

分 类 号：R739.4[医药卫生—肿瘤] R730.23[医药卫生—临床医学]