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

向伟.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[J].内科急危重症杂志,2019,25(1):58-63

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DOI：10.11768/nkjwzzzz20190119

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作者	单位	E-mail
向伟	武汉大学中南医院	huixiao@whu.edu.cn

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中文摘要:

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、2000μmol/L)作用24、48、72h，阿糖胞苷单用(0、10、50、100、200、400、800nmol/L)24h，普萘洛尔(400μmol/L)联合阿糖胞苷(10、50、100、200、400、800nmol/L)作用24h。普萘洛尔(0、200、400、800μmol/L)作用U937细胞24h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G0/G1期和减少G2/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G0/G1期细胞阻滞、损伤DNA，激活内、外源性凋亡途径有关。

英文摘要:

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, 800,1000,2000μmol/L) for 24, 48 and 72h, cytarabine alone (0,10,50,100,200,400,800nmol/L)and treated by cytarabine (0,10,50,100,200,400,800nmol/L) combined with propranolol ( 400μmol/L). Then, the U937 cells were treated by propranolol (0, 200, 400 and 800μmol/L) for 24h. 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. SPSS19. 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 G2/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.