The Effects Of Different HBx Expression On The Proliferation And Invasion Of HepG2 Hepatocellular Carcinoma Cells And Sorafenib Resistance

Objective: To investigate the effects of different expression levels of hepatitis B virus X(HBx)in HepG2 hepatocellular carcinoma(HCC)cells on proliferation,invasion,and sorafenib resistance.Methods: HepG2 cell lines were infected by lentiviral packaging of HBx-expressing plasmids and HBx-interfering fragments to construct stable HCC cells with different HBx expression levels.Among them,HepG2 cell line without treatment was the blank control group.HepG2 cell line infected with HBx high expression sequence lentivirus was the HBx high expression group and infected with lentivirus of HBx high expression negative control sequence was the negative control group.The interference group was infected with the HBx interference sequence virus based on the HBx high expression group to reduce HBx expression.Interference control group as interference group but with infected empty vector virus.The q PCR and western blotting was used to measure the protein expression of HBx in each group of HepG2 cells.The proliferation,invasion ability,and Sorafenib semi-inhibitory concentration(IC50)of HCC cells under different HBx expression levels were respectively detected by Cell proliferation assay kit,Transwell invasion assay,and Cell Titer-Glo(CTG)kit.Results: The result of qPCR and Western blotting showed that the stable expression cell lines of each group were successfully constructed.The cell proliferation ability of the HBx high expression group was better than that of the blank control and negative control groups,and the cell proliferation ability of the interference group was lower than that of the interference control and HBx high expression groups,and the differences were all statistically significant(P < 0.05).The number of cells crossing Matrigel gel was(46.2±4.1),(50.7±5.1)and(48.2±5.2)in the blank control group,negative control group,and interference group,respectively.The number of cells crossing Matrigel gel in the HBx high expression group(124.2±8.3)and the interference control group(117.2±7.5)were higher than the above three groups,respectively,and the differences were all statistically significant(P < 0.05).The IC50 of cells in the HBx high expression group and the interference control group were(5.36±0.31)μmol/L and(5.48±0.20)μmol/L,respectively,which were higher than those in the blank control group,the negative control group,and the interference group(4.75±0.22)μmol/L,(4.60±0.14)μmol/L and(3.98±0.03)μmol/L.The differences were all statistically significant(P < 0.05).Conclusion: HBx high expression promoted the proliferation and invasion of HepG2 HCC cells,enhanced the ability to sorafenib resistance,and inhibited apoptosis.