抑制低氧诱导因子-1α对PC12细胞存活的影响  

Effect of echinomycin on the cell viability of PC12 cells after it inhibits hypoxia inducible factor-1α

作　　者：李冉[1] 王勇[1] 宫晓丽[1] 范明[2] 王晓民[1] 朱玲玲[2] 汪璇[1] 

机构地区：[1]首都医科大学基础医学院生理学系,教育部神经变性病重点实验室,北京100069 [2]军事医学科学院基础医学研究所认知科学研究室,北京100850

出　　处：《首都医科大学学报》2012年第2期182-186,共5页Journal of Capital Medical University

基　　金：国家重点基础研究发展计划项目(973)(2011CB504100;2010CB944801);国家自然科学基金项目(30831160514);北京市自然科学基金项目(5102010)~~

摘　　要：目的通过抑制低氧诱导因子-1α转录活性,研究低氧诱导因子-1α在低氧诱导的细胞死亡中的作用。方法①将PC12细胞接种于12孔板,在20%常氧和3%低氧环境中培养24 h,在光镜下拍照并应用计数方法记录PC12细胞的生长情况。②将PC12细胞接种于96孔板,在常氧和低氧环境中培养24 h后,用细胞活力检测剂(a novel tetrazolium compound,3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt,MTS)检测PC12细胞的生长情况。③在常氧和低氧条件下分别用5 nmol/L、50 nmol/L、500 nmol/L的低氧诱导因子-1α抑制剂棘霉素处理PC12细胞24 h和48 h后,用MTS法检测细胞活力。结果①细胞计数和MTS法检测细胞活力的结果均显示3%的低氧条件可明显诱导PC12细胞死亡。②不同浓度的低氧诱导因子-1α抑制剂棘霉素处理PC12细胞24 h之后,低氧和常氧的细胞活力没有显著区别。③不同浓度的低氧诱导因子-1α抑制剂棘霉素处理PC12细胞48 h之后,低氧不再促进细胞死亡,相反,低氧条件处理的细胞活力明显好于常氧。结论低氧会促使PC12细胞死亡,棘霉素抑制HIF-1α的转录活性24 h后,低氧对PC12细胞的促死亡作用减弱;48 h后,低氧组的细胞活力还可超过常氧组。这表明,低氧下过量的HIF-1α转录激活对细胞是有害的。Objective To clarify the role of hypoxia inducible factor（HIF）-1α in the process of hypoxia-induced cell death through inhibiting its transcription activity.Methods ① PC12 cells were seeded in the 12-well plate and treated for 24 h in normoxia（20% O2） or hypoxia（3% O2）.Then photomicrographs under light microscope were taken and analyzed for situation of cell growth by cell-counting method.② PC12 cells were seeded in 96-well plate and treated for 24 h in 20% O2 or 3% O2.A test for cell survival called MTS was further used to detect the cell viability of PC12 cells.③ PC12 cells were treated with HIF-1α inhibitor-echinomycin for 24 h or 48 h at 5 nmol/L,50 nmol/L and 500 nmol/L,respectively.Then MTS was used to detect the cell viability.Results ① 3% O2 induced the death of PC12 cells significantly contrary to 20% O2.② Echinomycin administration eliminated the cell survival difference between 20% O2 and 3% O2 at 24 h.③ After a 48 h-treatment with echinomycin,it even reversed the hypoxia-enhanced cell death relative to normoxia.Conclusion Hypoxia could promote the death of PC 12 cells.After we used HIF-1α inhibitor echinomycin for 24 h,the death-promotion role of hypoxia was weakened.It even reversed the hypoxia-enhanced cell death after a 48 h-treatment with echinomycin.All these data suggested that the excessive transcription activation during hypoxia was detrimental to PC 12 cells.

关 键 词：低氧 低氧诱导因子1Α PC12细胞 棘霉素 

分 类 号：R32[医药卫生—人体解剖和组织胚胎学]