氯喹抑制的自噬对地西他滨促进髓性白血病细胞凋亡的影响

doi:10.13481/j.1671-587x.20170515

摘要/Abstract

摘要： 目的：研究氯喹与地西他滨联合应用对白血病K562和KG-1a1Aor1a细胞凋亡的影响，探讨自噬对地西他滨诱导白血病细胞凋亡的作用，阐明其作用机制。方法：体外培养髓性白血病K562和KG-1a1Aor1a细胞，分为空白对照组、地西他滨（10 μmol·L^-1）单用组和氯喹（50 μmol·L^-1）联用地西他滨组（联用组）。联用组细胞使用氯喹孵育6 h后再与其他组细胞同时开始实验。孵育24及48 h后，CCK-8法检测细胞数量并计算增殖抑制率，流式细胞术检测细胞凋亡率和线粒体膜电位。Q-PCR法检测Atg7及Atg12基因表达水平，Western blotting法检测LC3蛋白表达。结果：孵育24及48 h后，与空白对照组比较，地西他滨和联用组K562和KG-1a1Aor1a细胞数量数量明显减少（P < 0.05或P < 0.01）；凋亡率明显升高（P < 0.05或P < 0.01），线粒体膜电势明显增加（P < 0.05或P < 0.01）；与地西他滨组比较，联用组K562和KG-1a1Aor1a细胞明显减少，细胞数量凋亡率明显升高（P < 0.05）。孵育24 h后，与空白对照组比较，地西他滨组K562和KG-1a1Aor1a细胞Atg7、Atg12和LC3-Ⅱ/LC3-Ⅰ相对表达水平明显升高（P < 0.05或P < 0.01）；与地西他滨组比较，联用组K562和KG-1a1Aor1a细胞Atg7、Atg12和LC3-Ⅱ/LC3-Ⅰ相对表达水平明显降低（P < 0.05或P < 0.01）。结论：地西他滨具有促进白血病细胞凋亡的作用，而联用氯喹可以抑制自噬从而增强地西他滨诱导细胞凋亡的作用。

关键词: 地西他滨, 氯喹, 自噬, 细胞凋亡

Abstract: Objective: To study the influence of chloroquine combined with decitabine in the apoptosis of leukemia K562 cells and KG-1a1Aor1a cells, to explore the effect of autophagy on the leukemia cell apoptosis induced by decitabine, and to clarify its mechanism. Methods: The leukemia K562 and KG-1a1Aor1a cells were cultivated in vitro and divided into blank control group, decitabine group (10 μmol·L^-1) and chloroquine(50 μmol·L^-1) combined with decitabine group (combined group). The leukemia cells in combined group were pre-treated with chloroquine for 6 h before experiment. After treatment with drugs for 24 and 48 h, the number of cells was detected and by CCK-8 method the inhibitory rates of proliferation cells were calculated;the apoptotic rates and mitochondrial membrane potential were detected by flow cytometry. Q-PCR method was carried out to determine the gene expression levels of Atg7 and Atg12, and Western blotting was used to test the protein expression of LC3. Results: After treatment for 24 and 48 h, the number of K562 and KG-1a1Aor1a cells in decitabine group and combined group were decreased compared with blank control group(P<0.05 or P<0.01); the apoptotic rates and mitochondrial membrane potential were remarkably increased (P<0.05 or P<0.01). Compared with decitabine group, the number of K562 and KG-1a1Aor1a in combined group was significantly decreased, and the apoptotic rates were remarkably increased (P<0.05). After treatment for 24 h, the expression levels of Atg7, Atg12 and LC3-Ⅱ/LC3-Ⅰ in the leukemia K562 and KG-1a1Aor1a cells in decitabine group were significantly increased compared with blank control group (P<0.05 or P<0.01);the expression levels of Atg7, Atg12 and LC3-Ⅱ/LC3-Ⅰ in the leukemia K562 and KG-1a1Aor1a cells in combined group were significantly decreased compared with decitabine group (P<0.05 or P<0.01). Conclusion: Decitabine could promote the apoptosis of leukemia cells, and the inhibition of autophagy by chloroquine can promote the apoptosis induced by decitabine.

Key words: decitabine, chloroquine, autophagy, apoptosis

中图分类号:

R733.7

邢丽娜, 任金海, 王颖, 王福旭, 蔡圣鑫. 氯喹抑制的自噬对地西他滨促进髓性白血病细胞凋亡的影响[J]. 吉林大学学报(医学版), 2017, 43(05): 937-942.

XING Lina, REN Jinhai, WANG Ying, WANG Fuxu, CAI Shengxin. Influence of autophagy inhibited by chloroquine in apoptosisof myelogenous leukemia cells promoted by decitabine[J]. Journal of Jilin University Medicine Edition, 2017, 43(05): 937-942.

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