氯喹通过激活P38MAPK通路诱导人肝癌HepG2细胞凋亡

doi:10.3969/j.issn.1004-616x.2018.06.003

癌变·畸变·突变 ›› 2018, Vol. 30 ›› Issue (6): 430-434.doi: 10.3969/j.issn.1004-616x.2018.06.003

氯喹通过激活P38MAPK通路诱导人肝癌HepG2细胞凋亡

丁文评¹, 陆元元¹, 童文侠¹, 周蔼斌¹, 孔祥², 陈意红¹

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

收稿日期:2018-08-14 修回日期:2018-10-15 出版日期:2018-11-30 发布日期:2018-11-30
通讯作者: 陈意红,E-mail:871969345@qq.com E-mail:871969345@qq.com
作者简介:丁文评,E-mail:dingwenping99@163.com。
基金资助:
国家自然科学基金（81600645）；吴阶平医学科研基金（320.6750.17395）；皖南医学院中青年科研基金（WK2016F17）

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

DING Wenping¹, LU Yuanyuan¹, TONG Wenxia¹, ZHOU Aibin¹, KONG Xiang², CHEN Yihong¹

1. Department of Radiotherapy, Yijishan Hospital Affiliated to Wannan Medical College, Wuhu 241001;
2. Department of Endocrinology, Yijishan Hospital Affiliated to Wannan Medical College, Wuhu 241001, Anhui, China

Received:2018-08-14 Revised:2018-10-15 Online:2018-11-30 Published:2018-11-30

摘要/Abstract

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40 μmol/L）氯喹作用不同时间（24、48和72 h）后对细胞增殖的影响；用DAPI染色观察氯喹处理细胞24 h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；Western blot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、40 μmol/L氯喹（10 μmol/L氯喹作用24 h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、40 μmol/L氯喹处理24 h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、40 μmol/L氯喹能诱导HepG2细胞凋亡（P均 < 0.05）；Western blot结果显示20和40 μmol/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。

关键词: 氯喹, 肝癌, 凋亡, P38MAPK通路, P53

Abstract: 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 μmol/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 μmol/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 μmol/L led to a significant increase in nuclear concentration and shrinkage. Flow cytometry analyses showed that 10,20 and 40 μmol/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 μmol/L groups while Caspase-3 and P38MAPK did not change. Expression of Bcl-2 was decreased after treatment with 20 and 40 μmol/L. CONCLUSION: Chloroquine inhibited HepG2 cell growth and induced apoptosis. The underlying mechanism might involve the P38MAPK signaling pathway.

Key words: chloroquine, hepatocellular carcinoma, apoptosis, P38MAPK pathway, P53

中图分类号:

R730.1

丁文评, 陆元元, 童文侠, 周蔼斌, 孔祥, 陈意红. 氯喹通过激活P38MAPK通路诱导人肝癌HepG2细胞凋亡[J]. 癌变·畸变·突变, 2018, 30(6): 430-434.

DING Wenping, LU Yuanyuan, TONG Wenxia, ZHOU Aibin, KONG Xiang, CHEN Yihong. Induction of apoptosis in HepG2 hepatocellular carcinoma cells by chloroquine via the P38MAPK pathway[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(6): 430-434.

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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

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