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目的 研究不同程度缺氧条件下 ,肺癌A5 4 9细胞内过氧化物酶体增殖物激活受体α(PPARα)和缺氧诱导因子 1α(HIF 1α)的表达状况 ,及反义封闭HIF 1α后 ,对PPARα表达的影响 ,以了解PPARα和HIF 1α的相关性。方法 首先将A5 4 9细胞分为 4组 :正常对照组 (A组 )、缺氧 2 4小时组(B组 )、缺氧 4 8小时组 (C组 )、缺氧 72小时组 (D组 )。观察不同缺氧时间对PPARα及HIF 1α蛋白和mRNA表达水平的影响。为观察HIF 1α对PPARα表达状况的影响 ,再设计 4组 :对照组 2 (E组 ) ,HIF 1α反义寡核苷酸组 (F组 ) ,HIF 1α正义寡核苷酸组 (G组 ) ,HIF 1α错义寡核苷酸组 (H组 )。结果 正常对照组 ,PPARα和HIF 1α蛋白及mRNA均有少量表达 ;随着缺氧时间的延长 ,两者的表达水平逐渐升高。在缺氧 2 4小时 ,两者mRNA和蛋白开始增高 ,4 8小时明显增高 ,至 72小时达到高峰。反义封闭HIF 1α后 ,PPARα蛋白及mRNA的表达水平明显下降。结论 PPARα及HIF 1α的表达水平随肿瘤细胞缺氧信号的加强而增加 ,且PPARα受HIF 1α的调控。
Objective To investigate the expression of peroxisome proliferator - activated receptor α (PPARα) and hypoxia - inducible factor 1α (HIF 1α) in lung cancer A549 cells under hypoxia and the effects of antisense blocking HIF 1α On the expression of PPARα in order to understand the correlation between PPARα and HIF-1α. Methods A549 cells were divided into 4 groups: normal control group (group A), hypoxia 24 hours group (group B), hypoxia 48 hours group (group C), hypoxia 72 hours group (group D ). To observe the effect of different hypoxia time on PPARα and HIF 1α protein and mRNA expression levels. To observe the effect of HIF-1α on the expression of PPARα, four groups were designed: control group 2 (group E), HIF 1α antisense oligonucleotide group (group F), HIF 1α sense oligonucleotide group (group G) , HIF 1α missense oligonucleotide group (H group). Results The protein and mRNA of PPARα and HIF-1α were all slightly expressed in normal control group. With the prolongation of hypoxia time, the expression of both protein and mRNA increased gradually. At 24 hours of hypoxia, both mRNA and protein began to increase, 48 hours significantly increased to 72 hours to reach the peak. Antisense blocking HIF 1α, PPARα protein and mRNA expression levels decreased significantly. Conclusion The expression of PPARα and HIF-1α is increased with the increase of hypoxia signal in tumor cells, and PPARα is regulated by HIF-1α.