普萘洛尔对急性髓系细胞白血病U937细胞增殖和凋亡的影响  

Effect of propranolol on proliferation and apoptosis of acute myeloid leukemia U937 cells

作　　者：向伟[1] 肖晖[1] 宋燕燕[1] 肖慕然 XIANG Wei;XIAO Hui;SONG Yan-yan;XIAO Mu-ran(Zhonghan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院,武汉430071

出　　处：《内科急危重症杂志》2019年第1期58-63,共6页Journal of Critical Care In Internal Medicine

摘　　要：目的:探究普萘洛尔对急性髓系细胞白血病U937细胞的增殖和凋亡的影响及可能的机制。方法:采用CCK8法检测不同药物对U937细胞增殖率的影响,分别为普萘洛尔(0、200、400、800、1000、2 000μmol/L)作用24、48、72 h,阿糖胞苷单用(0、10、50、100、200、400、800 nmol/L)24 h,普萘洛尔(400μmol/L)联合阿糖胞苷(10、50、100、200、400、800 nmol/L)作用24 h。普萘洛尔(0、200、400、800μmol/L)作用U937细胞24 h后,用流式细胞术检测细胞的凋亡和周期、线粒体膜电位的变化,用碱性彗星实验检测DNA的损伤,Caspase-Glo试剂盒检测Caspase-3、8、9相对活性的变化,Western blot法检测Bcl-2、Bax蛋白表达量的变化。结果:普萘洛尔可以有效抑制U937细胞的增殖,其效应具有一定的时间(200、400μmol/L,P<0.05)和剂量相关性(0、200、400、800μmol/L,P<0.01),普萘洛尔还可协同阿糖胞苷的抗肿瘤效应。流式、彗星、Caspase、Western blot结果分别表明普萘洛尔可增加细胞凋亡率,增加G_0/G_1期和减少G_2/M期细胞比例,降低线粒体膜电位水平,诱导DNA损伤,增强Caspase3、8、9活性,上调Bax的表达、下调Bcl-2的表达,且在实验浓度范围内与剂量呈相关性(P<0.05)。结论:普萘洛尔可抑制急性髓系细胞白血病U937的增殖并诱导其凋亡,这一机制可能与其诱导G_0/G_1期细胞阻滞、损伤DNA,激活内、外源性凋亡途径有关。Objective:To observe the effect of propranolol on proliferation and apoptosis of human acute myeloid leukemia cell line U937,and explore the possible mechanisms.Methods:The CCK8 was used to detect the proliferation rate of U937 cells after treatment by propranolol at different concentrations(0,200,400,800,1 000,2 000μmol/L)for 24,48 and 72 h,cytarabine alone(0,10,50,100,200,400,800 nmol/L)and treated by cytarabine(0,10,50,100,200,400,800 nmol/L)combined with propranolol(400μmol/L).Then,the U937 cells were treated by propranolol(0,200,400 and 800μmol/L)for 24 h.We used a series of tests to detect the apoptosis rate,cell cycle,mitochondrial membrane potential,DNA damage and the change of the activity of Caspase-3,8,9,the expression of Bcl-2 and Bax.SPSS 19.0 was used to analyze the data.Results:CCK8 revealed that propranolol inhibited proliferation and cooperated with cytarabine in anti-proliferative effect on U937 cells and the effect has a certain time(200,400μmol/L,P<0.05)and dose correlation(0,200,400,800μmol/L,P<0.01).Flow cytometry showed that propranolol increased apoptosis rate and proportion in G0/G1 phase,decreased proportion in G2/M phase and mitochondrial membrane potential in U937 cells(P<0.05).The comet assay showed that propranolol induced DNA damage in a dose-dependent manner(P<0.05).Caspase-Glo assay showed propranolol enhanced the activity of Caspase-3,8,9.The expression of Bax was upregulated,and the expression of Bcl-2 was downregulated by Western blotting,and the effect is related to the dose in the experimental concentration range(P<0.05).Conclusion:The propranolol can inhibit the proliferation and induce the apoptosis of U937 cells,which was involved in arresting cell cycle in G0/G1 phase,damaging DNA,and activating endogenous and exogenous apoptotic pathways.

关 键 词：急性髓系细胞白血病 普萘洛尔 增殖 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]