Preliminary Study Of Wnt Pathway Regulator Pygo2 In The Pathogenesis Of Hepatocelluar Carcinoma

OBJECTS1.To investigate the expression of Wnt pathway regulartory factor Pygo2 in HCC,adjacant tissues and HCC cell lines;evaluate the prognostic value of Pygo2 in hepatocellular carcinoma.2.To explore the impact of knocking out Pygo2 on biological characteristics of cells such as viality and proliferation;preliminarily study the mechanism of Pygo2 played in the development of HCC by affecting Wnt/?-catenin signaling pathway targeted genes.3.To study the relationship between inflammatory cytokines IL-22 and Pygo2 and the application value of serum IL-22 levels in diagnosing the recurrence of hepatocellular carcinoma.METHODS1.Explorig the expression of Pygo2 in HCC and adjacent tissues by Real-time PCR,immunohistochemistry;using Western blot to detect the expression of Pygo2 in 5 different metastatic potential HCC cell lines;analyzing the correlation between Pygo2 expression and prognosis in patients with HCC.2.Establishing knockout SK-Hep1 and Hep3 B cell lines of Pygo2 by sh RNA,exploring the influence of knockout Pygo2 on cell viality and proliferation by CCK-8?clone formation,tumor-bearing of nude mice?immunohistochemistry?Real-time PCR?dual luciferase report gene and preliminarily studying the mechanism of Pygo2 played in the development of HCC by affecting Wnt/?-catenin signaling pathway targeted genes.RESULTS1.Pygo2 expression is higher in HCC than adjacent tissues at transcriptional and protein level(P <0.01);Pygo2 is positively correlated with invasion and metastasis potential of HCC cell lines;Pygo2 is negatively related to the prognosis of HCC patients.2.By experiments of CCK-8,clone formation,tumor-bearing of nude mice,we have found that the ability of vality,proliferation and tumor growth in HCC cell lines has decreased compared to the control;by dual luciferase report gene and Real-time PCR,we have found the expression of Wnt pathway targeted genes had decreased after knockout Pygo,especially the expression of c-myc;Pygo2 is positively correlated with c-myc at transcriptional level.3.The levels of serum IL-22 was positively correlated with the expression of Pygo2,and the levels of serum IL-22 in patients of recurrence was significantly highly than non-recurrence patients.CONCLUSIONS1.Pygo2 is highly expressed in HCC,it has potential value in HCC diagnosis and treatment.2.Pygo2 may involve in the development of HCC by affecting Wnt/?-catenin signaling pathway targeted genes.3.Serum IL-22 level is a independent prognostic factor in patients of HCC after curative resection.