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中国临床药理学与治疗学 ›› 2004, Vol. 9 ›› Issue (6): 695-697.

• 研究原著 • 上一篇    下一篇

重组人白血病抑制因子体外诱导HL-60细胞凋亡与bcl-2及p5表达的关系

杜冰, 朱道银1, 唐恩洁   

  1. 川北医学院分子生物学研究所, 组织胚胎学胚教研室, 南充637007, 四川;
    1重庆医科大学微生物学教研室, 重庆400016
  • 收稿日期:2004-02-14 修回日期:2004-03-20 发布日期:2020-11-22
  • 通讯作者: 唐恩洁,女, 教授, 硕士生导师, 主要从事分子生物学、免疫学和微生物学研究。Tel:0817-2242780
  • 作者简介:杜冰, 硕士。E-mail:n13990884328 @163.com
  • 基金资助:
    四川省教育厅重点基金项目(No01LA65)

Relation between recombinant human leukemia inhibitory factor(rh-LIF) induced apoptosis of HL-60 cells in vitro and expression of bcl-2 and p53

DU Bing, ZHU Dao-Yin1, TANG En-Jie   

  1. Institute of Molecular Biology, North Sichuan Medical College, Nanchong 637007, Sichuan, China;
    1Department of Microbiology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China
  • Received:2004-02-14 Revised:2004-03-20 Published:2020-11-22

摘要: 目的 探讨重组人白血病抑制因子(rh-LIF) 诱导HL-60 细胞凋亡的作用及其可能机制。 方法 不同剂量(10 ~ 4 000 U·ml-1) 的rh-LIF 作用HL-60 细胞3 d 后, 用流式细胞仪观察分析细胞周期变化并检测细胞凋亡率, 用TUNEL 法检测原位细胞凋亡情况;分别用免疫组化法和原位杂交法检测rh-LIF 作用2、4、6 d 后P53 蛋白及p53 mRNA、bcl-2 mRNA 表达水平的改变。 结果 rh-LIF (10 ~ 4 000 U·ml-1) 作用d 3, 细胞凋亡率较对照组显著增加, 其中以1 000 U·ml-1组的作用最强;rh-LIF (800 U·ml-1) 作用2 ~ 6 d,P53 蛋白和p53 mRNA 表达也同步增高,而bcl-2 mRNA 表达显著降低(P <0.01)。 结论 rh-LIF 能诱导HL-60 细胞凋亡, 该作用与P53 蛋白表达增加和bcl-2 表达降低有关。

关键词: rh-LIF, 凋亡, HL-60 细胞, bcl-2, p53

Abstract: AIM: To investigate the effect of recombinant human leukemia inhibitory factor (rh-LIF) on human myeloid leukemia cell line HL-60. METHODS: HL-60 cells were treated with various concentrations of rh-LIF (10-4 000 U·ml-1) for a few days, cell cycle and cell apoptosis rate were analysed by flow cytometry.Cell apoptosis was also observed under microscope with TUNEL.Expression of P53 protein after addition of rh-LIF (800 U ·ml-1) was observed by immunohistochemistry, and p53 mRNA and bcl-2 mRNA were assayed by in situ hybridization. RESULTS: Cell apoptosis rates (CAR) in rh-LIF groups by flow cytometry were higher than those in control group, and 1 000 U·ml-1 rh-LIF had the most potent effect on induction of apoptosis. There was also a marked increase of positive-cells by TUNEL method in a concentration-dependent pattern on the 3rd day after addition of rh-LIF (10-1 000 U · ml-1).Following addition of rh-LIF (800 U·ml-1) and incubation for 2 to 6 days, the expression of P53 protein and p53 mRNA of HL-60 cells increased obviously, and bcl-2mRNA expression decreased in a time-dependent pattern(vs control, P <0.01). CONCLUSION: rh-LIF can induce apoptosis, and the effects may be related to the increase of P53 protein and decrease of bcl-2 mRNA expression.

Key words: recombinant human leukemia inhibitory factor, apoptosis, HL-60 cell, bcl-2, p53

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