基础医学与临床 ›› 2008, Vol. 28 ›› Issue (8): 867-872.

• 研究论文 • 上一篇    下一篇

氯化钴化学模拟低氧对肿瘤细胞增殖及凋亡的影响

韩建群 余明华 戴旻 李宏伟 修瑞娟   

  1. 中国医学科学院微循环研究所 中国医学科学院微循环研究所 中国医学科学院微循环研究所 中国医学科学院微循环研究所
  • 收稿日期:2008-05-26 修回日期:2008-06-10 出版日期:2008-08-25 发布日期:2008-08-25
  • 通讯作者: 修瑞娟

Influence of cobalt chloride on the proliferation and apoptosis of tumor cells

Jian-qun HAN, Min-hua YU, Min DAI, Hong-wei LI, Rui-juan XIU   

  1. Institute of Microcirculation,CAMS & PUMC Institute of Microcirculation,CAMS & PUMC
  • Received:2008-05-26 Revised:2008-06-10 Online:2008-08-25 Published:2008-08-25
  • Contact: Rui-juan XIU

摘要: 目的 观察氯化钴(CoCl2)对体外培养的肿瘤细胞增殖和凋亡的影响,探讨合理的体外化学模拟低氧模式。方法 (1)MTT方法和流式细胞术检测不同浓度CoCl2在相应时间内对A549和HeLa细胞活力以及增殖和凋亡的影响。(2)免疫印迹测定HIF-1α和凋亡蛋白的表达。结果 (1)CoCl2(≤200?mol/L)在24h内对肿瘤细胞活力影响较轻,增加浓度或延长处理时间明显降低细胞活力。(2)CoCl2(200?mol/L)引起细胞早期凋亡。增加浓度(800?mol/L)导致晚期凋亡和坏死。(3)CoCl2(200?mol/L)诱导两种肿瘤细胞 HIF-1α上调,24h达高峰(P<0.05),增加浓度或延长处理时间则下调。在A549细胞CoCl2引起Bax /Bcl-2和P53表达上调;P53不参与CoCl2引起的HeLa细胞凋亡。结论 CoCl2对肿瘤细胞的增殖凋亡的影响呈时间和浓度依赖性,进行化学模拟低氧时,要考虑CoCl2的作用浓度和时间。

关键词: 氯化钴, A549, HeLa, 凋亡, 低氧诱导因子1-α

Abstract: Objective To investigate the effect of cobalt chloride on tumor cells proliferation and apoptosis in vitro, to explore the reasonable strategy of chemical hypoxia induced by CoCl2. Methods Two tumor cell lines (A549 and HeLa) were respectively exposed to CoCl2 (0.05~2mmol/L) for different time period (4~48h), cells viability、proliferation and apoptosis were maesured by MTT and FCM methods. HIF-1α and related apoptosis proteins expression were detected by Western blot. Results Cells viability were weakly changed in low concentration(≤200?mol/L)of CoCl2 within 24h. However, higher dose or prolonged challenge of CoCl2 significantly decreased cell survival rate (p<0.05). Most of the damaged cells were early apoptosis population after CoCl2 (200?mol/L) incubation within 24h, major of the damaged cells were late apoptosis and necrosis part after CoCl2(800?mol/L) treatment. CoCl2 (200?mol/L) exposure implicated in up-regulating the expression of HIF-1α, Bax, P53 and down-regulating the expression of Bcl-2 in A549 cell within 24h. However, higher concentration (~400?mol/L) or prolonged CoCl2 challenge lead to HIF-1α expression decreasing. Similar results were observed in HeLa cell, except P53. Conclusions Effect of CoCl2 on the tumor cells proliferation and apoptosis is dose and time-dependent. But prolonged or higher dose of cobalt exposure was not positive to expected chemical hypoxia effect (HIF-1α expression). The concentration and duration of CoCl2 administration should be taken into consideration in chemical hypoxia.

Key words: Cobalt chloride, A549 cell, HeLa cell, Apoptosis, HIF-1α