吉林大学学报(医学版)

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新型免疫抑制剂FTY720诱导K562细胞凋亡及其机制 

张宸豪，李妍，陈为，陈爽，方芳，赵良中   

  1. （吉林医药学院检验学院病原生物学教研室,吉林 吉林 132013）
  • 收稿日期:2014-08-01 出版日期:2014-11-28 发布日期:2015-01-18
  • 通讯作者: 李 妍 E-mail:（Tel:0431-64500461,E-mail:liyan@jlmpc.cn）
  • 作者简介:张宸豪（1972-），男，吉林省吉林市人，副教授，医学博士，主要从事抗感染免疫方面的研究 。
  • 基金资助:

    国家自然科学基金资助课题（82102953）；吉林省科技厅中青年科技创新领军人才及团队资助课题（20130521018JH）；吉林省科技厅重点科技攻关资助课题（20140203012YY）；吉林省教育厅科研基金资助课题（2012第487号，2012第420号）

Apoptosis of K562 cells  induced by a new immunosuppressive agent  FTY720 and its mechanism

ZHANG Chen-hao,LI Yan,CHEN  Wei,CHEN Shuang,FANG  Fang,ZHAO Liang-zhong   

  1. (Department of Pathogeny and Microbiology,Department of Medical Laboratory,Jilin Medical College,Jilin 132013,China)
  • Received:2014-08-01 Online:2014-11-28 Published:2015-01-18

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摘要：

目的：探讨新型免疫抑制剂FTY720诱导K562细胞凋亡的效应及其机制，为临床白血病的疗提供实验依据。方法：体外培养人白血病K562细胞，分为空白对照组和FTY720处理组，FTY720处理组细胞分别采用6 μmol•L-1 FTY720诱导3、6和12 h，或采用2、4、6和8 μmol•L-1 FTY720处理24 h。采用流式细胞术分析细胞凋亡百分率、活性氧（ROS）水平、线粒体膜电位（MMP）和细胞周期。采用WST-1 还原法检测FTY720作用下K562细胞增殖抑制率。结果：流式细胞术检测，与空白对照组比较，6 μmol?L-1 FTY720诱导细胞3、6和12 h后，细胞凋亡率随时间延长而升高（P<0.01）；与空白对照组比较，2、4、6和8 μmol•L-1 FTY720诱导细胞24 h 后，细胞凋亡率随药物浓度增加而升高（P<0.01）。与空白对照组比较，随FTY720浓度增加，K562细胞内ROS水平增加（P<0.01），同时其MMP下降（P<0.01）。细胞周期分析，与空白对照组比较，随FTY720浓度增加，G0/G1期细胞百分率增加（P<0.05），而S期和G2/M期细胞百分率减少（P<0.05）。WST-1 还原法分析，与空白对照组比较，FTY720处理72 h后，2、4、6和8 μmol?L-1FTY720处理组K562细胞增殖抑制率［（24.0±4.1）%、(46.4±3.9)%、(67.0±4.8)%和（88.2±5.6）%］明显升高（P<0.01），FTY720对K562细胞的半数抑制浓度（IC50）为5.5　μmol•L-1。结论：FTY720可通过导致细胞周期阻滞和激发ROS而诱导细胞凋亡来发挥抗肿瘤作用。

关键词: FTY720, 细胞凋亡, 活性氧, 细胞周期

Abstract:

Abstract:Objective  To study the  apoptosis of K562 cells induced by a new immunosuppressive agent FTY720 and its mechanism,and to provide experimental basis for the treatment of leukemia in clinic.Methods The K562 cells were cultured in vitro　and  divided into blank control  group and FTY720 treatment  group.The K562 cells in FTY720 treatment  group were treated with 6 μmol?L-1 FTY720  for 3,6,and 12 h,or treated with different concentrations of FTY720 (2，4，6,8 μmol•L-1 ) for 24 h.The apoptosis,level  of reactive oxygen species(ROS), mitochondrial membrane potential(MMP) and cell cycle were measured by flow cytometry.The  inhibitory rate of proliferation of  K562 cells after treated with FTY720 was detected by WST-1 reducting assay.Results The results of  flow cytometry showed that the percentages of apoptotic  cells were increased  after treated with 6 μmol•L-1    FTY720 for 3,6, and 12 h with the prolongation of time compared with blank control group（P<0.01）.The percentages of apoptotic cells were also increased  after treated with different concentrations of FTY720 for 24 h compared with blank control group（P<0.01）. Compared with blank control group,the ROS levels were  increased with the increasing of  FTY720 concentration,while the MMP was decreased（P<0.01）.Compared with blank control group,the percentage of cells at G0／G1 phase was increased,while those at S and G2/M phases were decreased with the increasing of  FTY720 concentration (P<0.05).The WST-1 reduction assay results indicated that the  inhibitory rates of proliferation of K562 cells after treated with 2,4,6, and 8 μmol•L-1  FTY720 for 72 h were (24.0±4.1)%，(46.4±3.9)%，(67.0±4.8)%, and (88.2±5.6)%, respectively,compared with blank control group.The concentration of FTY720 to result the   inhibitory rate of 50% (IC50) on K562 cells was 5.5 μmol•L-1 .Conclusion FTY720 could inhibit the proliferation of K562 cells   by blocking  cell cycle and  inducing  apoptosis through provoking ROS.

Key words: FTY720, apoptosis, reactive oxygen species, cell cycle

中图分类号: 

  • R392.12
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