甘草酸单铵盐调节急性早幼粒细胞白血病细胞NB4肿瘤干细胞样特性、氧化应激及线粒体的功能  

Effects of Monoammonium Glycyrrhizinate on Stem Cell-like Characteristics, Oxidative Stress and Mitochondrial Function of Acute Promyelocytic Leukemia NB4 Cells

作　　者：陈艳 温贤秀[1,3] 周晨燕 张熔[2] 吴炳菊 喻未[2] CHEN Yan;WEN Xian-Xiu;ZHOU Chen-Yan;ZHANG Rong;WU Bing-Ju;YU Wei(School of Nursing,Southwest Medical University,Luzhou 646000,Sichuan Province,China;Department of Pediatric,Sichuan Provincial Peopled Hospital,Sichuan Academy of Medical Sciences,Chengdu 610072,Sichuan Province,China;Nursing Department,Sichuan Provincial Peopled Hospital,Sichuan Academy of Medical Sciences,Chengdu 610072,Sichuan Province,China)

机构地区：[1]西南医科大学护理学院,四川泸州646000 [2]四川省医学科学院·四川省人民医院儿科,四川成都610072 [3]四川省医学科学院·四川省人民医院护理部,四川成都610072

出　　处：《中国实验血液学杂志》2022年第1期22-29,共8页Journal of Experimental Hematology

基　　金：四川省卫生和计划委员会科研课题(18PJ114)。

摘　　要：目的:探讨甘草酸单铵盐(MAG)对急性早幼粒细胞白血病细胞NB4肿瘤干细胞样特性、氧化应激及线粒体功能的影响。方法:CCK-8法检测急性早幼粒细胞白血病细胞NB4活力,筛选合适剂量;克隆法检测NB4细胞增殖;Western blot检测细胞周期相关蛋白表达;流式细胞术检测细胞凋亡并分选NB4干细胞阳性(CD133^(+));RT-PCR检测干细胞标志物(Oct4、ABCG2、Dclk1);荧光检测活性氧;试剂盒检测氧化应激标记物水平(MDA);流式细胞术检测线粒体膜电位变化;Western blot检测线粒体损伤指标相关蛋白的表达(Bax/BCL-2)。结果:与对照组相比,MAG浓度<5μmol/L时,细胞NB4活力不具有显著性差异;浓度>5μmol/L时,对细胞NB4生长的抑制作用明显增加(P<0.05),根据CCK-8实验结果,按浓度设置MAG 0、5、10和20μmol/L;与对照组(MAG 0μmol/L)相比,除MAG 5μmol/L组差异无统计学意义外,NB4细胞增殖率、周期蛋白表达量、线粒体膜电位及干细胞CD133^(+)比例、标记物m RNA水平(Oct4、ABCG2、Dclk1)均显著降低(P<0.05),NB4细胞凋亡率、活性氧水平、MDA含量及Bax/BCL-2表达均显著增加(P<0.05)。结论:MAG对急性早幼粒细胞白血病细胞NB4的影响具有剂量依赖性,随浓度的升高,抑制作用增强。其抗肿瘤机制可能与下调肿瘤干细胞样特性、促进氧化应激与破坏线粒体功能等有关。Objective:To investigate the effect of monoammonium glycyrrhizinate on the stem cell-like characteristics,oxidative stress and mitochondrial function of acute promyelocytic leukemia cells NB4.Methods:CCK-8 method was used to detect the viability of acute promyelocytic leukemia cells NB4,and the appropriate dose was screened;Cloning method was used to detect the proliferation rate of NB4 cell;Western blot was used to detect the expression of cell cyclerelated protein;flow cytometry was used to detect cell apoptosis and sort NB4 stem cells positive(CD133^(+));Stem cell markers(Oct4,ABCG2,Dclk1) were detected by RT-PCR;ROS was detected by fluorescence;The kit was used to detect the level of oxidative stress markers(MDA);The flow cytometry was used to detect the change of mitochondrial membrane potential;Western blot was used to detect the expression of mitochondrial damage index-related proteins(Bax/BCL-2).Results:Compared with the control group,if the concentration of MAG was less than 5 μ mol/L,the cell NB4 viability showed no significant difference;if the concentration was higher than 5 μ mol/L,the inhibitory effect on the growth of cell NB4 increased and showed significant difference(P<0.05),according to the results of CCK-8 experiment,four groups were set based on the concentration of MAG 0 μ mol/L,MAG 5 μ mol/L,MAG 10 μ mol/L,and MAG 20μ mol/L;compared with the control group(MAG 0 μ mol/L),the cells in MAG 5 μ mol/L group showed no significant difference,while the proliferation rate,cyclin expression,mitochondrial membrane potential,stem cell CD 133^(+)ratio,and marker mRNA level(Oct4,ABCG2,Dclk1) of NB4 cell were significantly reduced(P<0.05);the apoptosis rate,reactive oxygen species,MDA content and Bax/BCL-2 expression of NB4 cell significantly increased(P<0.05).Conclusion:Monoammonium glycyrrhizinate has a significant inhibitory effect on acute promyelocytic leukemia cells NB4,which may be related to the regulation of stem cell-like characteristics,oxidative stress and mitochondrial function.

关 键 词：急性早幼粒细胞白血病 甘草酸单铵盐 NB4 线粒体 氧化应激 CD133^(+) 

分 类 号：R733.71[医药卫生—肿瘤]