Construction Of Lentiviral Vector For TWEAK Gene And Its Expression And Function Study In Hepatoma Cells

Objective: Lentivirus vector of human tumor necrosis factor weak apoptosis inducers(TWEAK) gene was constructed and transfected into human hepatoma cell line Hep G2 to study the influences of TWEAK on Hep G2 cells in some associated genes expression level and biological characteristics.Methods: The entire coding region of TWEAK gene was amplified from p ORF5-h TWEAK plasmid by PCR to construct the Plentilox3.7-TWEAK plasmid. The recombinant plasmid vectors were packed into 293 T cells to product lentiviruses to transfect Hep G2 cells. q RT-PCR, western-blot and flow cytometry were used to measure the expression of TWEAK, Fn14, p38 MAPK and p-p38 MAPK and the change of cell proliferation, cell cycle, apoptosis and metastasis in TWEAK-Hep G2 cells.Results The entire coding region of TWEAK gene was amplified and cloned into Plentilox3.7. The lentiviruses produced by 293 T were efficient to transfect Hep G2 cells. In the Hep G2 cells transfected with TWEAK gene, the expression of TWEAK and Fn14 were significantly up-regulated, while p38 MAPK-mediated signal pathway was not activated. Meanwhile, cell proliferation, cell cycle, cell apoptosis and cell metastasis of Hep G2 transfected with TWEAK gene did not change obviously.Conclusions In this study, the recombinant inducible lentiviral vector containing TWEAK gene was successfully constructed and transfected into Hep G2 cells to overexpress TWEAK. It is preliminary indicated that TWEAK expression in Hep G2 cells can effectively induce Fn14 expression. However, the biological function needs further research.