国际肿瘤学杂志

国际肿瘤学杂志 ›› 2014, Vol. 41 ›› Issue (5): 371-375.doi: 10.3760/cma.j.issn.1673-422X.2014.05.016

• 论著 • 上一篇    下一篇

蛇床子素诱导HL-60细胞凋亡及其分子机制研究

王岩,姜国胜,任霞,黄宁,毕可红   

  1. 250062 济南大学 山东省医学科学院医学与生命科学学院（王岩）；山东省医学科学院基础医学研究所（姜国胜、任霞）；山东省千佛山医院血液科（黄宁、毕可红）
  • 收稿日期:2014-01-13 修回日期:2014-03-05 出版日期:2014-05-08 发布日期:2014-04-28
  • 通讯作者: 毕可红 E-mail:bikehong11@163.com
  • 基金资助:

    国家自然科学基金（81172792）

Induction of apoptosis by osthole in HL-60 cells and the molecular mechanism research

Wang Yan, Jiang Guosheng, Ren Xia, Huang Ning, Bi Kehong   

  1. School of Medicine and Life Sciences, University of Ji′nan-Shandong Academy of Medical Sciences， Ji′nan 250062 , China
  • Received:2014-01-13 Revised:2014-03-05 Online:2014-05-08 Published:2014-04-28
  • Contact: Bi Kehong E-mail:bikehong11@163.com

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摘要： 目的 研究蛇床子素对急性髓系白血病HL-60细胞增殖、凋亡的影响，并探讨其作用的分子机制。方法 CCK8实验检测蛇床子素对HL-60细胞的增殖抑制作用，Hoechst染色观察药物8 h作用后细胞的形态学变化，流式细胞术检测细胞凋亡情况，反转录聚合酶链式反应（RT-PCR）检测HL-60细胞的Bcl-2、Bax mRNA表达变化，Western bolt检测HL60细胞的活性caspase-3、caspase-8、caspase-9、Fas、FasL蛋白表达变化。结果 蛇床子素可明显抑制HL-60细胞增殖并诱导其发生凋亡，抑制率最高达（90.7±4.5）%，F=138.46,P=0.000;总凋亡率为33.6%，F=27.75,P=0.006。诱导后Bcl-2 mRNA表达下调，Bax mRNA表达上调，Bax/Bcl-2比值升高（F=210.12，P=0.000），活性caspase-3、caspase-8、caspase-9、Fas、FasL蛋白表达水平随药物作用时间延长而升高。结论 蛇床子素可抑制HL60细胞增殖并诱导凋亡，其诱导凋亡机制与同时激活线粒体途径和死亡受体途径相关。

关键词: Osthole, Leukemia, Apoptosis

Abstract: Objective To detect the effect of osthole on proliferation and apoptosis of HL-60 cells and its molecular mechanism. Methods HL-60 cells proliferation was measured through the CCK8 assay method. The cell morphological changes were observed by Hoechst33342 staining after 8 h of drug effect. Induction of apoptosis was determined by flow cytometry and fluorescent microscopy. Expressions of Bcl-2 and Bax mRNA were evaluated by RT-PCR, and the expressions of cleaved caspase-3, caspase-8, caspase-9, Fas and FasL were evaluated  by using western bolt assay. Results Osthole could inhibit the proliferation of HL-60 cells, the maximum inhibiting rate was （90.7±4.5）%， F=138.46, P=0.000; the apoptosis rate was 33.6%， F=27.75, P=0.006. The changes of apoptosis of cells and nucleus were shown in cell morphological observation. Osthole affected the decrease of the mRNA levels of Bcl-2 and the increase of the Bax mRNA levels via a dosedependent manner（F=210.12, P=0.000）. Western blotting demonstrated that osthole could lead to the increase of the expression levels of cleaved caspase-3, caspase-8, caspase-9, Fas and FasL in the HL-60 cell line via a timedependent manner. Conclusion Data suggests that osthole inhibits proliferation and induces apoptosis of HL-60 cells through mitochondriadependent pathway and deathreceptor pathway.

Key words: Osthole, Leukemia, Apoptosis