Expression of HIF-1α and VEGF in Glioma A172 Cells After Gene Intervention

Abstract

Objective To observe the changes of hypoxia induction factor-a(HIF-α) and vascular endothelial growth factor(VEGF) expression after in vitro gene intervention, so as to explore the role of HIF-1α in the regulation of angiogenesis in glioma. Methods CoCl2 chemical hypoxia method simulated tumor hypoxia environment, and reverse transcription polymerase chain reaction (qPCR), immunofluorescence method, Western blot method and ELISA double antibody sandwich method were used to compare the expression levels of related mRNA and protein in glioma A172 cells before and after gene intervention of HIF-1α. Results HIF-1α mRNA expression was not significantly changed under hypoxia, and VEGF mRNA expression was significantly increased. The protein expressions of HIF-1α and VEGF were increased successively in hypoxic culture at 8, 16 and 24 hours. After successful gene intervention in hypoxia culture for 24 hours, HIF-1α and VEGF protein expressions were significantly reduced. Compared with the untransfected group, the inhibition rates of transfection were 86.3% and 78.9% respectively, and the inhibition rate of VEGF mRNA expression was 81.5%. Conclusion VEGF expression is significantly reduced after HIF-1α expression is inhibited by siRNA intervention. The protein expression of HIF-1α in A172 cells is increased in the hypoxic environment, and HIF-1α promotes angiogenesis in glioma under hypoxia through transcriptional activation of VEGF.

Keywords

• glioma
• hif-α
• vegf
• genetic intervention