西罗莫司对人急性早幼粒白血病细胞HL60增殖的抑制作用  

Inhibition of Rapamycin on Proliferation of Human Acute Promyelocytic Leukemia HL60 Cells

作　　者：侯毅鞠[1] 常青[2] 国巍[3] 袁忠海[1] 李艳[1] 

机构地区：[1]吉林医药学院血液检验教研室,吉林吉林132013 [2]中国人民解放军总医院消化科,北京100853 [3]吉林大学第一临床医院肿瘤中心,长春130021

出　　处：《中国药学杂志》2013年第23期2002-2005,共4页Chinese Pharmaceutical Journal

基　　金：国家重大科技专项(2012ZX091013-301-003);吉林省教育厅科学技术研究项目(2012489)

摘　　要：目的探讨西罗莫司对人急性早幼粒白血病细胞HL60增殖、自噬、凋亡的影响。方法四甲基偶氮唑蓝(MTT)法观察西罗莫司对白血病细胞HL60的体外生长抑制作用,蛋白质印迹法(Western-blot)检测西罗莫司处理HL60细胞后自噬相关蛋白LC3-Ⅱ的表达情况,流式细胞术(FCM)检测西罗莫司干预后HL60细胞的周期分布,并通过DNA琼脂糖凝胶电泳检测凋亡。结果四甲基偶氮唑蓝法结果显示,1、5、10、20、40nmol·L^(-1)的西罗莫司能够抑制HL60细胞增殖,并呈浓度依赖性(r=0.97,P<0.05)。蛋白质印迹法结果显示,各浓度组LC3-Ⅱ表达水平高于对照组(P<0.01),具有促进细胞自噬作用;FCM显示西罗莫司干预后的白血病细胞株出现细胞周期阻滞,G_0/G_1期细胞比例逐渐增加,S期细胞明显减少,与对照组比较具有显著性差异(P<0.05);琼脂糖凝胶电泳未见凋亡细胞形成DNA片段。结论西罗莫司具有抑制人急性早幼粒白血病细胞HL60增殖的作用,在限定浓度范围内以诱导HL60细胞发生自噬为主而不是凋亡。OBJECTIVE To investigate the effect of rapamycin on proliferation, autophagy and apoptosis of human acute promye- locytic leukemia HL60 cells. METHODS The inhibitory effect of rapamycin on HL60 cell was detected by MTT assay. The expres- sions of LC3-II autophagy-related protein were determined by Western blot. The cell cycles were analyzed by flow cytometry （ FCM ）. Apoptosis were detected by DNA agarose gel electrophoresis. RESULTS After treatment with rapamycin of 0,1,5,10,20,40 nmol ~ L ~ for 24 h, proliferation of HL60 cells were inhibited in a dose-dependent manner （ r = 0. 97, P 〈 0. 05 ）. Western-blot shows that the expression of LC3-II protein of each concentration group was higher than that of the control group （ P 〈 0.01 ）. Rapamycin has a role in promoting cell autophagy. Compared with control group, more cells were arrested at G0/GI phases （P 〈 0. 05 ） and fewer cells were at S phases （P 〈 0. 05 ）. Agarose gel electrophoresis shows no apoptosis DNA fragments. CONCLUSION In given concentra- tion range,rapamycin can inhibit the proliferation of human acute promyelocytic leukemia HL60 cells by promoting autophagy rather than inducing apoptosis.

关 键 词：西罗莫司 急性早幼粒白血病 增殖 HL-60细胞 

分 类 号：R965[医药卫生—药理学]