喹乙醇诱导HepG2细胞自噬的实验研究  

Autophagic process induced by olaquindox in HepG2 cells

作　　者：赵东旭[1] 张朝明[1] 汤树生[1] 王丛丛[1] 张燊[1] 肖希龙[1] 

机构地区：[1]中国农业大学动物医学院药理和毒理教研室,北京100193

出　　处：《癌变．畸变．突变》2013年第3期168-172,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：中央高校基本科研业务费专项资金(2011JS009)

摘　　要：目的:研究喹乙醇对HepG2细胞自噬的影响。方法:分别用不同浓度(0、200、400、800μg/mL)喹乙醇染毒HepG2细胞24h后,进行单丹磺酰尸胺(monodansylcadaverine,MDC)染色,分别通过荧光显微镜和流式细胞仪检测细胞自噬发生情况。另外,分别用不同浓度喹乙醇(0、200、400、800μg/mL)染毒HepG2细胞24h和400μg/mL喹乙醇染毒HepG2细胞不同时间(0、1、3、6、12、24h)后,采用Westernblot方法检测自噬相关蛋白LC3-Ⅱ和Beclin1的表达。结果:随着喹乙醇染毒剂量增加,MDC阳性细胞的荧光强度及细胞内MDC荧光颗粒数目明显增加。流式结果显示,与对照组相比,200、400和800μg/mL染毒组MDC阳性细胞百分比均显著增加(P<0.05或<0.01)。随着喹乙醇染毒浓度和时间的增加,LC3-和Beclin1表达量均明显增加,其中400μg/mL喹乙醇染毒细胞24h组与对照组相比,LC3-Ⅱ和Beclin1的表达量均明显上调(P<0.01)。结论:喹乙醇诱导了HepG2细胞的自噬反应。OBJECTIVE： To investigate the effect of olaquindox on autophagy in human hepatoma G2 （HepG2） cells. METHODS：The HepG2 cells were treated with different concentrations （0,200,400,800 μg/mL） of olaquindox for 24 h. Then autophagy was analyzed by fluorescence microscope and flow cytometer following staining with monodansylcadaverine （MDC）. HepG2 cells were treated with different concentrations （0,200,400,800 μg/mL） of olaquindox for 24 h or with 400 μg/mL olaquindox for different periods （0,1,3,6,12,24 h）. Then the expressions of LC3-Ⅱ and Beclin 1 were determined by Western blot. RESULTS：Olaquindox-treated cells exhibited higher fluorescent density and more MDC-labeled particles in HepG2 cells compared with the control group. The percentage of MDC-positive cells was obviously increased after treatment with different concentrations （200,400,800 μg/mL） of olaquindox for 24 h. （P0.05 or P0.01）. The expressions of LC3-Ⅱ and Beclin 1 increased with different concentrations and different time points of olaquindox treatment. Moreover,compared with the control group,the expression of LC3-Ⅱ and Beclin 1 were increased significantly with treatment of 400 μg/mL olaquindox for 24 h （P0.01）. CONCLUSION： Olaquindox enhanced the autophagic process in HepG2 cells.

关 键 词：喹乙醇 自噬 MDC LC3-II BECLIN1 

分 类 号：R994.6[医药卫生—毒理学]