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

发布时间：2018-03-01 21:15

  本文关键词： 地西他滨 氯喹 自噬 细胞凋亡　出处：《吉林大学学报(医学版)》2017年05期 　论文类型：期刊论文

【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(10μmol·L-1)单用组和氯喹(50μmol·L-1)联用地西他滨组(联用组)。联用组细胞使用氯喹孵育6h后再与其他组细胞同时开始实验。孵育24及48h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48h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(P0.05或P0.01);凋亡率明显升高(P0.05或P0.01),线粒体膜电势明显增加(P0.05或P0.01);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(P0.05)。孵育24h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(P0.05或P0.01);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(P0.05或P0.01)。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。
[Abstract]:Aim: to study the effect of chloroquine combined with dicitabine on apoptosis of leukemia K562 and KG-1a1Aor1a cells, and to explore the effect of autophagy on the apoptosis of leukemic cells induced by dicitabine. Methods: K562 and KG-1a1Aor1a cells of myeloid leukemia were cultured in vitro and divided into blank control group. Dietabine (10 渭 mol 路L -1) and chloroquine (50 渭 mol 路L -1) combined with dietabine (combination group) were incubated with chloroquine for 6 h and then began the experiment simultaneously with other groups. After 24 and 48 h incubation, the number of cells was measured by CCK-8 method and the inhibitory rate of proliferation was calculated. Apoptosis rate and expression level of Atg7 and Atg12 gene were detected by flow cytometry and mitochondrial membrane potential. Results: after incubation for 24 and 48 hours, the expression of LC3 protein was detected by Western blotting. The number of K562 and KG-1a1Aor1a cells in the combination group decreased significantly (P0.05 or P0.01), the apoptosis rate increased significantly (P0.05 or P0.01), the mitochondrial membrane potential increased significantly (P0.05 or P0.01), and compared with the combination group, the number of K562 and KG-1a1Aor1a cells decreased significantly in the combination group. After incubation for 24 hours, compared with the control group, the relative expression levels of Atg7N / Atg12 and LC3- 鈪，

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