氯喹通过激活P38MAPK通路诱导人肝癌HepG2细胞凋亡  

Induction of apoptosis in HepG2 hepatocellular carcinoma cells by chloroquine via the P38MAPK pathway

作　　者：丁文评 陆元元[1] 童文侠[1] 周蔼斌 孔祥[2] 陈意红[1] DING Wenping;LU Yuanyuan;TONG Wenxia;ZHOU Aibin;KONG Xiang;CHEN Yihong(Department of Radiotherapy,Yijishan Hospital Affiliated to Wannan Medical College,Wuhu 241001;Department of Endocrinology,Yijishan Hospital Affiliated to Wannan Medical College,Wuhu 241001,Anhui,China)

机构地区：[1]皖南医学院弋矶山医院放疗科,安徽芜湖241001 [2]皖南医学院弋矶山医院内分泌科,安徽芜湖241001

出　　处：《癌变．畸变．突变》2018年第6期430-434,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(81600645);吴阶平医学科研基金(320.675017395);皖南医学院中青年科研基金(WK2016F17)

摘　　要：目的:研究氯喹对人肝癌HepG2细胞增殖和凋亡的影响,并初步研究其可能的机制。方法:体外培养HepG2细胞,采用MTT法检测不同浓度(0、10、20和40μmol/L)氯喹作用不同时间(24、48和72h)后对细胞增殖的影响;用DAPI染色观察氯喹处理细胞24h后细胞核的变化;流式细胞术检测氯喹对HepG2细胞凋亡的影响;Westernblot技术检测氯喹对HepG2细胞中Caspase-3、Bcl-2、Bax、P-P53、P38MAPK和P-P38MAPK蛋白表达的影响。结果:与对照组比较,在上述3个时间点10、20、40μmol/L氯喹(10μmol/L氯喹作用24h组除外)对人肝癌HepG2细胞的增殖均有明显抑制作用(P均<0.05);荧光显微镜下发现,10、20、40μmol/L氯喹处理24h后均可引起HepG2细胞不同程度的核浓集、固缩等典型凋亡形态变化;流式细胞术结果提示10、20、40μmol/L氯喹能诱导HepG2细胞凋亡(P均<0.05);Westernblot结果显示20和40μmol/L氯喹作用HepG2细胞后,P-P53、P-P38MAPK、剪切后活化型Caspase-3和Bax蛋白表达量增加(P均<0.05),Bcl-2表达下降,Caspase-3和P38MAPK表达无明显变化(P>0.05)。结论:氯喹能够抑制人肝癌HepG2细胞的生长并诱导其凋亡,其机制可能与P38MAPK通路有关。OBJECTIVE:To investigate effects of chloroquine on cell proliferation and apoptosis of a hepatocellular carcinoma cell line,HepG2.METHODS:Cultured HepG2 cells were treated with various concentrations of chloroquine(0,10,20 and 40 mol/L).Cell viability was determined by the MTT assay.Morphological changes in cell nuclei was detected using fluorescence microscope at 24 h after treatment.Cell apoptosis was detected by flow cytometry.Expression levels of Caspase-3,Bcl-2,Bax,P-P53,P38MAPK and P-P38MAPK were performed using Western blot analyses.RESULTS:After treatment with 10-40 mol/L chloroquine for 24,48 and 72 h,proliferation of HepG2 cells was significantly inhibited compared with the control group(P<0.05).In addition,treatment with 10,20 and 40 mol/L led to a significant increase in nuclear concentration and shrinkage.Flow cytometry analyses showed that 10,20 and 40 mol/L treatment significantly induced apoptosis(P<0.05).Increased in protein expression of P-P53,P-P38MAPK,cleaved Caspase-3 and Bax was detected in the 20 and 40 mol/L groups while Caspase-3 and P38MAPK did not change.Expression of Bcl-2 was decreased after treatment with 20 and 40 mol/L.CONCLUSION:Chloroquine inhibited HepG2 cell growth and induced apoptosis.The underlying mechanism might involve the P38MAPK signaling pathway.

关 键 词：氯喹 肝癌 凋亡 P38MAPK通路 P53 

分 类 号：R730.1[医药卫生—肿瘤]