Mechanism Research on Death of Hepatic Carcinoma Cells Induced by Chloroquine
- VernacularTitle:氯喹在诱导肝癌细胞死亡中的作用机制研究
- Author:
Zhenbin NG DI
1
;
Yinghong HI S
;
Yuanfei PENG
;
Kang SONG
;
Jian ZHOU
;
Jia FAN
Author Information
1. 复旦大学附属中山医院肝外科
- Keywords:
Autophagy;
Hepatocellular carcinoma;
Apoptosis;
Chloroquine
- From:
Chinese Journal of Clinical Medicine
2015;(2):135-139
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the role of chloroquine (CQ) during inducing the death of hepatic carcinoma cells and to explore the clinical application of autophagy‐related agents .Methods:Three kinds of hepatic carcinoma cell lines were chosen and treated with CQ at different concentrations for 24 hours .Then MTT assay was applied to determine the proliferation ,and prodium iodide (PI) stalning assay was used to detect the cell death .The specific apoptosis was examined by Hochest 33342 stalning , TUNEL stalning and Western blotting (caspase‐9 ) . The autophagic function was explored by GFP‐LC3 redistribution ,Western blotting (LC3) ,and electron microscopy .Results:MTT assay revealed that the proliferation of hepatic carcinoma cells was significantly inhibited by using CQ for 24 hours .The PI‐positive cells increased significantly after the treatment with CQ (≥ 20 μmol/L) .Hochest33342 stalning assay showed karyopyknosis and karyorrhexis .Meanwhile , TUNEL positive cells significantly increased and caspase‐9 was activated .However ,there was no significant change in the number of PI‐positive cells after the CQ treatment at concentrations of 5 μmol/L and 10 μmol/L .CQ treatment resulted in significant increase of GFP‐LC3‐positive cells and expression of LC3‐II .When the caspase inhibitor Z‐VAD‐FMK and 3‐MA or Atg5 siRNA were applied at the same time , 40 μmol/L CQ‐induced M HCC97‐L cell death was significantly inhibited . Conclusions:CQ can inhibit the autophagic degradation of hepatic carcinoma cells , and resulted in the accumulation of autophagosomes and autophagic proteins .High‐dose CQ directly induces the death of hepatic carcinoma cells ,and the death has both characteristics of apoptosis and autophagic death .
