低氧微环境对胎鼠成纤维细胞HIF-1α和PTEN表达及细胞周期的影响  

The effect of hypoxia on PTEN and HIF-1α expressions and cell cycle in MEFs

作　　者：赵丹[1] 唐嘉茵[1] 王艳艳[1] 刘慧雯[1] 

机构地区：[1]哈尔滨医科大学组织学与胚胎学教研室,哈尔滨黑龙江150081

出　　处：《解剖科学进展》2012年第4期305-309,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金资助项目(No.30971537);黑龙江省科技厅海外学人项目(LC02C13);哈尔滨市科技局项目(2002AA9CS151-05)

摘　　要：目的探讨低氧条件下HIF-1α和PTEN在胎鼠成纤维(MEFs)细胞增殖、分化中可能的调控作用,以及低氧对细胞周期的影响。方法低氧条件(5%O_295%N_2)下培养MEFs不同时间(0、2、4、6、8h)后,应用免疫细胞化学、RT-PCR检测方法,观察MEFs中HIF-1α和PTEN的表达并检测其细胞周期。结果免疫组化结果表明,HIF-1α及PTEN蛋白的表达均随低氧时间增长而逐渐增强。RT-PCR结果显示,HIF-1αmRNA吸光度值常氧组为(0.93±0.06),低氧2～4h组达高峰分别为(1.46±0.05)和(2.30±0.05),随后开始下降;PTENmRNA吸光度值常氧组为(0.78±0.08),低氧2～6h组逐渐达到高峰分别(1.10±0.11)和(1.61±0.23),8h开始呈现下降趋势(<0.05)。流式细胞仪分析显示,随着低氧时间延长,G0/G1期细胞从(86.72±0.41)%上升至(92.41±1.10)%,低氧使MEFs细胞周期阻滞于G1/G0期。结论低氧微环境阻滞MEFs细胞周期于G0/G1期可能与PTEN和HIF-1α过表达相关。Objective To study the effect of hypoxia on PTEN and HIF-1α expressions and cell cycle in mouse embryo fibroblasts（MEFs）. Methods MEFs were cultured at different times（0, 2, 4, 6, 8 h）under hypoxic conditions （5% 02 95% N2）. the expressions of HIF-1 α and PTEN in MEFs were observed using immunoeytochemistry and RT-PCR, and the cell cycle was monitored simultaneously. Results Immunohistoehemistry showed that the expression levels of HIF- 1 α and PTEN protein increased gradually with the increased hypoxic time. RT-PCR showed that absorbanee values of HIF-lα mRNA and PTEN mRNA products were much lower in normoxia group than in hypoxia, reached the peak in hypoxia 2-4h group （P〈0.05）, then decreased gradually. Flow cytometry analysis showed that the number of G0/G1 phase cells increased from （86.72 ± 0.41 ）% to （92.41 ± 1.10）% with prolonged hypoxia, cell cycle of MEFs stoped at G1/G0 phase. Conclusion The inhibition of hypoxia cell micro-environment on MEFs cells in G0/G1 phase might be related to the overexpression of HIF-1 α and PTEN.

关 键 词：低氧 PTEN HIF-1Α 细胞周期 胎鼠成纤维细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]