癌变·畸变·突变 ›› 2009, Vol. 21 ›› Issue (2): 109-113.doi: 10.3969/j.issn.1004-616x.2009.02.008

• 论著 • 上一篇    下一篇

白藜芦醇诱导HL-60细胞凋亡机制的实验研究

何顺华,李冠武,黄宁宇   

  1. 汕头大学医学院肿瘤分子生物学开放实验室,广东 汕头 515041
  • 收稿日期:2008-11-06 修回日期:2008-12-05 出版日期:2009-03-30 发布日期:2009-03-30

Molecular Mechanism of Resveratrol-induced Apoptosis in HL-60 Cells

HE Shun-hua, LI Guan-wu, HUANG Ning-yu   

  1. Open Laboratory for Tumor Molecular Biology, Medical College of Shantou University, Shantou 515041, Guangdong, China
  • Received:2008-11-06 Revised:2008-12-05 Online:2009-03-30 Published:2009-03-30

摘要: 背景与目的: 探讨白藜芦醇(resveratrol,Res)诱导人早幼粒白血病HL-60细胞凋亡的机制。 材料与方法: 用不同浓度Res作用HL-60细胞,倒置显微镜观察细胞的形态学变化,MTT法检测Res对HL-60细胞增殖的抑制作用,确定Res作用于HL-60细胞的半数抑制浓度(IC50)。Western blot 法检测Res作用HL-60后Caspase-3 活性的变化及GADD45α、Annexin A1、Bcl-2/Bax、Cleaved-Caspase 3表达的变化。 结果: Res处理后,HL-60细胞体积变小,细胞的折光性减弱,细胞内出现颗粒样物质,且随着Res浓度的增加上述形态变化增强。12.5~200 μmol/L浓度的Res可明显抑制HL-60细胞的增殖,具有时间和剂量依赖性(P<0.05),24 h的IC50为75.64 μmol/L。Res处理后,HL-60细胞Caspase-3 活性增强,12 h达高峰,24 h开始下降;细胞的Annexin A1、GADD45α、Bax、Cleaved-Caspase 3蛋白表达量升高,Bcl-2蛋白表达量下降,Bcl-2/Bax比值下降。 结论: Res抑制HL-60细胞增殖,并通过依赖Caspase-3途径诱导HL-60细胞凋亡,GADD45α、Annexin A1参与了凋亡过程。

关键词: 白藜芦醇, HL-60细胞, 凋亡, GADD45α, Annexin A1

Abstract: BACKGROUND AND AIM: To explore the molecular mechanisms of resveratrol in inducing apoptosis in human promyelocytic leukemia HL-60 cells. MATERIALS AND METHODS: After treatment with resveratrol at different concentrations for different times, the morphological changes of HL-60 cells were examined by phase contrast microscopy. The proliferation of HL-60 cells and the IC50 were analyzed by methyl thiazolyltetrazolium (MTT)assay.Caspase-3 acticity and expression levels of GADD45α,Annexin A1,Cleaved-caspase 3,Bcl-2 and Bax proteins in HL-60 cell were measured by Caspase-3 activity assay kit and Western blot. RESULTS: After treatment with resveratrol, the HL-60 cells became smaller,less refractive with the appearance of granular materials. The changes were associated with the increase of resveratrol concentration. After treatment with 12.5-200 μmol/L Res, HL-60 cell proliferation was markedly inhibited in a dose and time -dependent manner(P<0.05) , and the IC50 was 76.54 μmol/L. After treatment with Res for 0,12,24,48 h,the Caspase-3 activity in HL-60 cells peaked at 12 h,and then decreased. The expressions of Annexin A1,GADD45α,Bax,Cleaved-Caspase 3 were up-regulated and Bcl-2 was down-regulated in HL-60 cells by resveratrol. CONCLUSION: Resveratrol could inhibit the proliferation and induce apoptosis of HL-60 cells, the effect might be associated with Caspase-3 activity. Expressions of GADD45α and Annexin A1 participated in the apoptotic process modulated by resveratrol.

Key words: resveratrol, HL-60, apoptosis, GADD45α, Annexin A1