雷公藤甲素对U251细胞自噬活性的影响及机制研究  

The influence and mechanism of Triptolide on autophagy in human glioma cell line U251

作　　者：惠双[1] 郭宏强 王博[1] 王媛[1] 张成辉[1] 

机构地区：[1]河南省南阳市中心医院肿瘤内科,473000 [2]郑州大学附属肿瘤医院肿瘤内科,郑州450000

出　　处：《重庆医学》2016年第23期3179-3181,3184,共4页Chongqing medicine

基　　金：国家自然科学基金青年基金(81101797)

摘　　要：目的探讨雷公藤甲素对U251细胞自噬活性的影响及机制。方法采用四甲基偶氮唑蓝(MTT)法检测雷公藤甲素对U251细胞的增殖抑制率、半数抑制浓度(IC50);免疫荧光化学法测定细胞中LC3-Ⅱ表达,Western blot法测定LC3-Ⅱ/LC3-Ⅰ、Beclin-1、磷酸化P70、磷酸化Erk、磷酸化AKT蛋白的表达。结果雷公藤甲素作用于U251细胞后,细胞增殖受抑制,24h的IC50为0.73μmol/L,48h的IC50为0.27μmol/L。免疫荧光法测定6、12、24h后LC3-Ⅱ水平增高;Western blot法测定LC3-Ⅱ水平、LC3-Ⅱ/LC3-Ⅰ比值随时间增高;Beclin-1水平增高,磷酸化P70、磷酸化Erk表达降低、磷酸化AKT无明显变化。结论雷公藤甲素可抑制U251细胞增殖并促进自噬,并可通过上调Beclin-1表达和抑制mTOR的活性来促进自噬。Objective . To explore the variation of autophagic activity in U251 ceils after treatment with Triptolide and its mechanisms. Methods We measured the proliferation inhibition rate, half inhibitory concentration （ICso） by MTT assay; LC3-Ⅱ protein expression were tested by immunofluorescence assay; protein expression of LC3-Ⅱ/LC3-Ⅰ , Beclin-1, Phospho-P70, Phos- pho-Erk and Phospho-AKT were measures by Western bIot. Results After treatment with Triptolide, the proliferation of U251 cells was inhibited,with the IC50 of 0.73 μmol/L at 24 h and the IC50 of 0.27 μmol/L at 48 h;the expression of LC3-Ⅱ were in- creased at 6,12,24 h;the expression of LC3-Ⅱ , LC3-Ⅱ/LC3-1 and Beclin-1 increased, the expression of Phospho-P70 and Phos- pho-Erk decreased and the expression of Phospho-AKT had no obvious change. Conclusion Triptolide could inhibit the prolifera- tion of U251 cells and induce cell autophagy;Triptolide could increase the autophagy by upregulating Beclin-1 expression and inhibi- ting the activity of mTOR.

关 键 词：雷公藤内酯 自噬 胶质母细胞瘤 U251细胞 

分 类 号：R285[医药卫生—中药学]