Wisp-1: A Novel Mediator Linking HBV And Hepatocellular Carcinoma

Objective: Observe the changes of Wnt-induced-secreted-protei-1 ( WISP-1) in quantity caused by the HBX protein with the methods of HIC and Western blot, and try to explain the mechanism by which could the HBV make the Hepatocellula Carcinoma(HCC) come into being in a new angle. Methods: (1) Collected samples of tissues from HCC, the paratumor of HCC and normal livers. Tested the expression of WISP-1 in these samples and compared the disparities. (2) Transfect plasmid of pc-DNA3.1(+) and plasmid of pc-DNA3.1(+)-HBX into HepG2 cells by Lipofectamine2000. Obtain the stable strains named G0 and Gx after a long-term screening by G418. (3) Tested the expression of WISP-1 in mRNA level in G0 and Gx using PCR and compared the disparities. (4) Tested the expression of WISP-1 in protein level in G0 and Gx using Western blot and compared the disparities. Results: (1)WISP-1 positive rate: HCC group > the paratumor group> normal liver group. The result was confirmed by X2 test and the p value< 0.01. (2) The stable strains of Gx which could secrete HBX protein continuously and of G0 which was used as the negative control of Gx were got after a month of screening of G418. (3) The result of Western blot : HBX was positive in Gx and negative in G0, which showed the transfection was successful. (4) The result of PCR: the expression of WISP-1 in Gx was higher than that in G0 on the level of mRNA. (5) The result of Western blot: the expression of WISP-1 in Gx was higher than that in G0 on the level of protein. Conclusion: WISP-1 was highly expressed in HCC. The HBV could up-regulate the expression of WISP-1 by its X protein when it had invaded into the hepatocell and thus promoted the development of HCC.