胃癌BGC-823细胞中HIF-1α、HIF-2α对PKM2的调控作用

Regulation effect of HIF-1α and HIF-2α on PKM2 in BGC-823 cells of gastric cancer

目的研究siRNA特异性沉默人胃癌BGC-823细胞中缺氧诱导因子(HIF-1α、HIF-2α)后PKM2的表达情况,探索HIF-1α、HIF-2α对胃癌细胞内PKM2的调控作用。方法用Realtime RT-PCR及Western blot法检测转染HIF-1α、HIF-2α及Control siRNA后细胞内PKM2表达情况。结果 RNA干扰技术能有效沉默BGC-823细胞中的HIF-1α、HIF-2α。转染HIF-1α、HIF-2αsiRNA后,PKM2在mRNA及蛋白表达水平均下降(P<0.05),其中siRNA抑制HIF-1α后,PKM2下调最明显;转染Control siRNA后,PKM2在mRNA及蛋白表达水平均无明显变化(P>0.05)。结论 HIF-1α和HIF-2α均参与调控PKM2的表达,其中HIF-1α起主要调控作用,HIF-1α、HIF-2α与PKM2共同促进肿瘤的Warburg效应。

Objective To compare the expression of PKM2 in BGC-823 cells after transfection with small in- terfering RNA of hypoxia-inducible factors （HIF-1α and HIF-2α ） and investigate the effect of HIF-1α and HIF-2α on the regulation of PKM2 in gastric cancer. Methods Small RNA interference （RNAi） was used to silence BGC-823 cells. The mRNA and protein expression levels of HIF-1α , HIF-2α and PKM2 were detected by quantitative real-time reverse transcription PCR and western blotting. Results Small RNA interference （RNAi） could effectively silence the expressions of HIF-1α and HIF-2α genes and down-regulate the expression of PKM2 （P 〈 0. 05 ）, and the levels of PKM2 decreased most obviously when being transfected with HIF-lct siRNA. The expression of PKM2 levels did not change significantly （ P 〉 0.05 ） when being transfected with control siRNA. Conclusion The results suggest that HIF-1α and HIF-2α are both involved in regulating PKM2. Compared to HIF-2α pathway, HIF-1α pathway plays a major role in regulating PKM2 ,and thus they all contribute to the Warburg effect of tumor.