中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (27): 5062-5066.doi: 10.3969/j.issn.1673-8225.2010.27.028

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

冬凌草乙素对白血病HL-60细胞的增殖抑制作用

刘晓丹1，刘文达2，王春芝1，徐  妍1，林东军1，刘培庆3，黄河清3，吴传斌3，肖若芝1，黄仁魏1，刘加军1   

  1. 中山大学附属第三医院，1血液科，2输血科，广东省广州市 510630；3中山大学药学院药理学与毒理学实验室，广东省广州市 510080
  • 出版日期:2010-07-02 发布日期:2010-07-02
  • 通讯作者: 刘加军，副教授，硕士生导师，中山大学附属第三医院血液科，广东省广州市 510630 jiajun.l@163.com
  • 作者简介:刘晓丹★，男，1981年生，中山大学附属第三医院血液病学专业在读硕士，主要从事抗白血病药物对血液肿瘤细胞的凋亡机制研究。lenovo381@126.com
  • 基金资助:

    国家自然科学基金项目(30772782)。教育部新世纪优秀人才支持计划资助项目( NCET-0721)。

Anti-proliferation effect of Ponicidin on leukemia HL-60 cells

Liu Xiao-dan1, Liu Wen-da2, Wang Chun-zhi1, Xu Yan1, Lin Dong-jun1, Liu Pei-qing3, Huang He-qing3, Wu Chuan-bin3, Xiao Ruo-zhi1,Huang Ren-wei1, Liu Jia-jun1   

  1. 1 Department of Hematology, 2 Department of Transfusion, Third Hospital of Sun Yat-sen University, Guangzhou  510630, Guangdong Province, China; 3 Laboratory of Pharmacology and Toxicology, School of Pharmaceutical Science, Sun Yat-sen University, Guangzhou  510080, Guangdong Province, China
  • Online:2010-07-02 Published:2010-07-02
  • Contact: Liu Jia-jun, Associate professor, Master’s supervisor, Department of Hematology, Third Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China jiajun.l@163.com
  • About author:Liu Xiao-dan★, Studying for master’s degree, Department of Hematology, Third Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China lenovo381@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30772782*; the Outstanding Talent Support Project of Ministry of Education of China, No. NCET-0721*

PDF

436

被引次数

13

摘要：

背景：已有研究证明，冬凌草乙素可以通过诱导许多实体肿瘤的细胞凋亡而发挥抗肿瘤作用，目前，冬凌草乙素对白血病HL-60细胞的作用尚未见资料报道。
目的：观察冬凌草乙素对白血病HL-60 细胞的体外增殖抑制作用及其机制。
方法:以不同浓度的冬凌草乙素(10~50 μmol/L)作用于体外培养的HL-60 细胞24，48，72 h，应用MTT法检测细胞生长抑制率，流式细胞术检测细胞周期，免疫印迹法检测Caspase-3及其裂解底物多聚(ADP-核糖)聚合酶(PARP)表达水平的变化，并对细胞周期调节蛋白P21、P16的表达水平进行检测。
结果与结论：冬凌草乙素可显著抑制细胞的生长及诱导细胞发生凋亡，呈现出明显的量-效与时-效关系。流式细胞术检测结果表明细胞主要被阻滞在G0/G1期，50 μmol/L的冬凌草乙素作用48 h后可以出现典型的亚G1期峰(细胞凋亡峰)。免疫印迹法检测结果显示Mr32 000的Caspse-3酶被活化出现Mr20 000亚单位片段，同时Caspse-3的底物PARP被裂解出现Mr89 000的亚单位片段，免疫印迹法检测结果还显示50 μmol/L的冬凌草乙素作用不同时间后，细胞周期调节蛋白P21及P16的表达水平逐渐升高。结果提示冬凌草乙素在体外对HL-60 细胞具有显著的细胞周期阻滞作用，并诱导细胞发生凋亡，通过上调细胞周期调节蛋白P21及P16的表达水平及激活Caspse-3可能是冬凌草乙素引起HL-60细胞G0/G1阻滞及诱导细胞发生凋亡的重要作用机制之一。

关键词: 冬凌草素, HL-60 细胞, 细胞凋亡, 白血病, 作用机制

Abstract:

BACKGROUND: Recent studies have shown that Ponicidin (PON) plays a significant role in anti-cancer effect via inducing apoptosis in various human cancer cell lines. At present, few reports have addressed the PON effects on leukemia cell lines HL-60 cells.
OBJECTIVE: To investigate the anti-proliferation effect of PON on leukemic HL-60 cells in vitro and its mechanisms of action.
METHODS: HL-60 cells in culture medium in vitro were given different concentrations of PON (10-50 μmol/L) for 24, 48 and 72 hours. The inhibitory rates were measured by MTT assay, and cell cycle was detected by flow cytometry. Expressions of Caspase-3 and poly(ADP-ribose) polymerase (PARP) as well as cell cycle modulator P16 and P21 were detected by Western blot assay.
RESULTS AND CONCLUSION: PON could remarkably inhibit the growth of HL-60 cells and cause apoptosis; the suppression was in time- and dose-dependent manners. Flow cytometry detection revealed that the cells were mainly arrested in G0/G1 phase and the subG1 cells were also found clearly in the histogram following 50 μmol/L PON treatment. Western blot results showed cleavage of the Caspase-3 zymogen protein (Mr32 000), with the appearance of its Mr 20 000 subunit, and a cleaved Mr 89 000 fragment of PARP after the cells were treated by different concentrations of PON. Western blot analysis also revealed that P21 and P16 expression was gradually up-regulated following 50 μmol/L PON treatment. Results have indicated that PON demonstrates in vitro anti-leukemia effect in HL-60 cells mainly associated with cell cycle arrest and apoptosis. Up-regulation of the expression of P16 and P21 as well as activation of Caspase-3 may be one of its most important mechanisms of inducing HL-60 cell inhibition in G0/G1 phase or cell apoptosis.

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