二甲双胍对急性单核细胞白血病细胞株THP-1增殖、分化和凋亡的影响  

Effect of Metformin on Proliferation,Differentiation and Apoptosis of THP-1 Cells

作　　者：张电安[1] 孟凡静[1] 周俊[1] 马萍[2] 宋旭光[1] 曹江[1] 

机构地区：[1]徐州医学院附属医院血液科 [2]徐州医学院医学技术学院,江苏徐州221002

出　　处：《中国实验血液学杂志》2015年第3期633-636,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金(81470303);江苏省"六大人才高峰"资助项目(2011-WS-067)

摘　　要：目的:探讨二甲双胍(metformin)对体外培养的急性单核细胞白血病细胞株THP-1细胞增殖、分化和凋亡的影响,并分析其可能的作用机制。方法:用不同浓度的二甲双胍分别对THP-1细胞进行体外干预,24 h、48 h后用CCK-8法检测细胞的增殖情况;20 mmol/L二甲双胍干预24 h后用Annexin V/7-AAD双标记法分析细胞凋亡情况;流式细胞术检测细胞表面分化抗原CD11b、CD14的变化;实时定量PCR检测BCL-XL、BAX、BIM、Caspase-3mRNA表达的变化。结果:CCK-8法检测显示二甲双胍对THP-1细胞增殖具有显著的抑制作用,且作用呈时间和剂量依赖性;20 mmol/L二甲双胍干预THP-1细胞24 h后,流式细胞术检测结果显示CD11b、CD14阳性细胞率无明显变化(P>0.05);Annexin V/7-AAD标记的流式细胞术检测显示,实验组和对照组早期凋亡细胞比例分别为(2.02±0.85)%和(4.46±1.33)%,晚期凋亡细胞比例分别为(1.43±0.83)%和(3.31±0.59)%,在实验组明显高于对照组(P<0.05);20 mmol/L二甲双胍诱导THP-1细胞过程中BCL-XL、BIM表达无明显变化,BAX、Caspase-3表达显著增加(P<0.01)。结论:二甲双胍能有效抑制THP-1细胞增殖并促进细胞凋亡,但对THP-1细胞分化无明显影响,其促进凋亡机制可能与上调BAX及Caspase-3基因有关。Objective：To investigate the effect of metformin on proliferation, differentiation and apoptosis of THP-1 cells and explore its possible mechanism. Mehods：THP-1 cells were cultured with different concentrations of metformin for 24 h and 48 h. The cell proliferation was evaluated by CCK-8, the cell apoptosis was analyzed by Annexin V/7-AAD double labeling, the expression of CD14 and CDllb （surface differentiation antigens on THP-1 cells） was evaluated by flow cytometry, the BCL-XL, BAX, BIM and caspase-3 mRNA expressions of THP-1 cells were detected by real time quantitative PCR. Results： Metformin could significantly inhibit the growth of THP-1 cells in a time- and dosedependent manner. After treated with 20 mmol/L metformin for 24 h, the expressions of CD14 and CDllb in THP-1 cells didn＇t change much（ P 〉0.05 ）, the early apoptosis rates in exprimental and control groups were （2.02 ± 0.85 ）% and （4.46 ± 1.33 ）% respectively, the late apoptosis rates in experimental and control groups were （1.43 ± 0.83 ）% and （3.31 ± 0.59）% respectively. In process of inducing effect of 20 mmol/L metformin on THP-1 cells, the expressions of BCL-XL and BIM did not significantly changed, while the expressions of BAX and caspase-3 significantly increased （ P 〈 0.01 ）. Conclusion： Metformin can effectively inhibit proliferation and induce apoptosis of THP-1 cells. However, it has no significant effect on differentiation of THP-1 cells, its mechanism inducing apoptosis maybe related with up-regulating BAX and caspase-3.

关 键 词：二甲双胍 白血病 细胞凋亡 THP-1细胞 

分 类 号：R733.71[医药卫生—肿瘤]