Mechanism Study Of Hepatitis B Virus X Protein Regulating Gankyrin In Hepatocarcinogenesis

Objective To investigate the role of HBX protein in the expression ofGankyrin, trying to clarify the possible molecular mechanism ofpathogenesis of HBV-related HCC by regulating the expression of coreprotein β-catenin in HBX protein and Wnt/β-catenin signal pathway.Methods1The expression of Gankyrin in HepG2and HepG2.2.15at thecellular level was observed by RT-PCR and Western-Blot respectively at themRNA and protein levels;2HepG2and SMMC-7721cells were transfectedwith HBX protein adenovirus vector, the expression of Gankyrin wasobserved by RT-PCR and Western-Blot respectively at the mRNA andprotein levels;3HpeG2.2.15cells were transfected with SiHBX moleculeswith LipofectamineTM2000, the expression of Gankyrin was detected byRT-PCR and Western-Blot respectively at the mRNA and protein levels;4. β-catenin adenovirus vector transfected into HepG2, SMMC-7721cells,detect the expression of β-catenin and Gankyrin by Westernblot;5.Overexpression HBX in β-catenin Silenced HepG2, SMMC-7721cells detect the expression of β-catenin and Gankyrin by western blot.Results1.The level of Gankyrin expression in HepG2cells was higherthan that in HepG22.15cells, indicating the regulation of Gankyrinexpression by some HBV genome encoded proteins;2.Compared to thecontrol group,the expression level of Gankyrin in experimental group was significantly increased with the overexpression of HBX protein in2.HepG2and SMMC-7721cells, indicating that HBV genome encoded HBX proteinhas up-regulated the expression of Gankryin;3.Compared to the controlgroup,the expression level of Gankyrin in experimental group wassignificantly reduced in the absence of HBX protein in HepG2.2.15cell,indicating that HBV genome encoded HBX protein could regulate theexpression of Gankyrin directly;4Compared to the control group, theexpression level of Gankyrin in experimental group was dramaticallydecreased, after interfering of Wnt/β-catenin signaling pathway coreprotein β-catenin in the overexpression of HBX HepG2and SMMC-7721cells.Which has confirmed that the role of HBX protein in the expression ofGankyrin through Wnt/β-catenin signaling pathway.Conclusion HBX-Wnt/β-catenin–Gankyrin signal transductionpathways are exist in Hepatocarcinogenesis.Inhibiting the function ofβ-catenin can interrupt the process in HBX upregulating the expressin ofGankyrin, It may provide a new treatment method for HCC.