The Study Of Dynamic Changes Of Notch Signaling Patyway In The Development Of HBx-HCC In Vivo

Objective: To construct a mosuse model carrying hepatitis B virus X stably for a long time,and to explore the mechanism of HBx causing hepatocelluar carcinoma in vivo.Methods: Hepatic progenitor cells transfected with HBx were injected into KM mice via hepatic portal vein to construct animal models.The liver tissues were harvested at 30 d,90d,180 d,and 360 d after operation.Western blot and q RT-PCR were used to detect the expression of HBx,the histopathological sections was observed by HE staining;Western blot and q RT-PCR to detect the dynamic expression of Notch1,Notch4,Hes1 at each time point;then intraperitoneal injection of the inhibitor DAPT at180 d,Western blot and q RT-PCR to detect the changes in the expression of apoptosis factors Bcl-2 and Bax and cell-cycle-related factors CDK4,Cyclin E and Cyclin D1.Results: The animal model was successfully constructed,HBx is persistently expressed in mice and liver cancer occurred at 360 d.Compared with the control group,the m RNA and protein levels of Notch1,Notch4,and Hes1 were significantly up-regulated at each time point of HBx-EGFP-14-19 groups;the expression of anti-apoptotics factor Bcl-2was significantly up-regulated in the HBx group,while pro-apoptotics factor Bax was down-regulated,the protein and m RNA expression of CDK4,Cyclin E,Cyclin D1 were increased in the HBx group.And the inhibitor DAPT group significantly reversed the above trend.Conclusion: HBx causes anti-apoptosis and abnormal cell-cycle-accelerating in mice by persistently activating the Notch pathway,which ultimately leads to HCC.