The Role Of HIF-2?-VEGF-Notch In Residual Hepatocellular Carcinoma After High Intensity Focused Ultrasound

Objective: To explore the expression of HIF-1?,HIF-2?,VEGF and Notch in residual hepatocellular carcinoma after high intensity focused ultrasound(HIFU).Methods: The best Subline were established and expanded that survived from HIFU treatment.The expression of VEGF was detected by immunofluorescence staining in treated and control group.Real-time PCR and Western blotting were used to measure the expression of HIF-1?,HIF-2? and VEGF mRNA and protein at 0h,6h,12 h and 24 h after hypoxia treatment in both treated and control groups.Then,the treated group was infected with LV-shRNA-VEGF LV-sh RNA-HIF-2? and LV-sh RNA-NC,and the expression of VEGF and HIF-2?,Notch,DLL4 mRNA and protein were quantified by real-time PCR and Western blotting.Results: The VEGF protein expressed in green fluorescence was mainly expressed in the cytoplasm and expressed both in the treated and control groups.The expression of HIF-1? in the treated and control groups reached peak at the 12 th hour after hypoxic treatment and then decreased,while the expression of HIF-2? and VEGF increased with the time past.(P <0.05).The expression of VEGF,Notch and DLL4 in VEGF modulated group was lower than that in the treated group(P <0.05).The expression of HIF-2? and VEGF in HIF-2? modulated group was lower than that in the treated group(P <0.05).The expression of HIF-2?,VEGF,Notch and DLL4 in the treated group were significantly higher than control group(P <0.05),but were similar to negative groups(P> 0.05).Conclusion: HIF-2?-VEGF-Notch signal pathway is involved in the regulation of angiogenesis in residual hepatocellular carcinoma after high intensity focused ultrasound,which further reveals the new mechanism of rapid development of HIFU residual hepatocellular carcinoma.