【摘要】 目的通过RNA干扰技术沉默A549细胞中乏氧诱导因子-1α(HIF-1α)的表达,初步探讨HIF-1在TGF-β介导的血管生成素样基因4(ANGPTL4)表达上调过程中的作用。方法转染了HIF-1α-shRNA的A549细胞(实验组)、转染空质粒的A549细胞(对照组)以及未转染的A549细胞(空白组)分别给予不同时间的TGF-β1干预,采用RT-PCR检测各组细胞中的HIF-1α与ANGPTL4的mRNA水平,Westernblot检测各组细胞中HIF-1α与ANGPTL4蛋白的表达变化。结果 TGF-β1处理0、12、24、48 h后,相比于对照及空白组,实验组中HIF-1α与ANGPTL4的mRNA及蛋白表达水平显著降低(P<0.05),且对照组及空白组中HIF-1α与ANGPTL4的mRNA及蛋白水平均与TGF-β1干预具有显著的时效相关性(P<0.05)。而实验组内HIF-1α与ANGPTL4未观察到上述变化趋势。结论 HIF-1可能是ANGPTL4的上游调控基因,并且参与TGF-β-Smad信号通路介导的ANGPTL4表达上调。更多还原

【Abstract】 Objective To discuss the effect of HIF-1α on TGF-β mediated upregulation of ANGPTL4 through down regulating the expression of HIF-1α in A549 cell lines by RNA interference.Method HIF-1α-shRNA transfected A549 cells(expand discuss the effect of HIF-1 in TGF-β mediated upregulation of ANGPTL4,exprimental group),empty plasmid transfected A549 cells(negative control group) and A549 cells(blank group) were treated with TGF-β1 for various time interval.RT-PCR and Western blot were used to determine the mRNA and protein levels of HIF-1α and ANGPTL4 in the three groups,respectively.Results After TGF-β1 treatment for 0,12,24 or 48h,the mRNA and protein levels of HIF-1α and ANGPTL4 were significantly reduced in comparison with that in negative control and blank groups(P<0.05).Additionally,the mRNA and protein levels of HIF-1α and ANGPTL4 were correlated with action time of TGF-β1 in a time-effect manner in negative control and blank groups(P<0.05).However,the similar changes that regarding to HIF-1 and ANGPTL4 were not observed in experimental groups.Conclusion The probable role of HIF-1 as an upstream regulator of ANGPTL4,may be involved in regulating the TGF-β-Smad pathway mediated overexpression of ANGPTL4.更多还原