The Influence Of Genistein On The Expression Of HIF-1α And VEGF In HUVEC-12 Cells Induced By Hypoxia

Objective: To research the influence of Genistein on the expression of HIF-1αand VEGF in HUVEC-12 cells induced by CoCl2 that is hypoxia-simulating agent.Methods: (1)HUVEC-12 cells were treated with CoCl2 at different concentration (0μM,25μM,50μM,100μM,150μM,200μM)for 8h to explore the concentration -effect of CoCl2 on cellular growth and with 100μM CoCl2 for different time (0h,1h,2h,4h,8h,12h,24h) to test the time -effect . The relationship between the expression of HIF-1αand VEGF was detected by CHX;(2)Pretreated with different concentrations of Genistein(0μM,50μM,100μM,150μM,200μM)for half an hour, HUVEC-12 cells were cultured with 100μM CoCl2 for 8h. MTT assay was adopted to detect the effect of HUVEC-12 cells proliferation; the expression of HIF-1αand VEGF mRNA were detected by RT-PCR; the expression of HIF-1αand VEGF protein were detected by Western Blot.Results: RT-PCR showed that CoCl2 have no effect on the expression of HIF-1αmRNA in different concentration or for different time. But it can respectively increase the expression of VEGF mRNA with a peak at 100μM CoCl2 in a concentration dependent manner,and for 8 hours in a time-dependent manner, the expression of HIF-1αand VEGF protein is significantly up regulated by CoCl2 exposure. CHX as the inhibitor of HIF-1αdecreased the expression of HIF-1αand VEGF protein. MTT showed that proliferation of HUVEC-12 cells was increased significantly by CoCl2 simulated hypoxia with a peak at 100μM CoCl2; Genistein inhibited the expression of VEGF mRNA in a concentration-dependent manner. But have no effect on the expression of HIF-1αmRNA. Western Blot show Genistein can inhibit the expression of HIF-1αand VEGF protein induced by CoCl2 exposure. And Genistein can inhibit the proliferation of HUVEC-12 cells induced by CoCl2 in a concentration-dependent manner.Conclusion: (1) CoCl2 can increase the expression of VEGF mRNA and HIF-1αand VEGF protein; CoCl2 can increase the proliferation of HUVEC-12 cells.(2). Genistein can inhibit the expression of HIF-1αand VEGF induced by CoCl2; Genistein can inhibit the proliferation of HUVEC-12 cells induced by CoCl2.