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AIM: To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells. METHODS: HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by Iipid-mediated transfection, including transient and stable transfection. Positive clones were screened by incubating in the selective medium with 600 μg/mL G418 and named HL-7702/HBV-encoded X protein (HBx) cells. The expressions of Fas/FasL, Bax/Bcl-2, and c-myc mRNA were measured by semi-quantitative RT-PCR in HL-7702/HBx and control group, respectively. RESULTS: RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. By semi-quantitative RT-PCR analysis, Bax and c-myc mRNA levels in HL-7702/HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702/HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702/HBx cells of transient and stable transfection were significantly lower than those in control. CONCLUSION: HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas/FasL, Bax/Bcl-2, and c-myc gene in a dose-dependent manner.
METHODS: HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by Iipid-mediated transfection, including transient and Stable transfection. Positive clones were screened by incubating in the selective medium with 600 μg / mL G418 and named HL-7702 / HBV-encoded X protein (HBx) cells. The expressions of Fas / FasL, Bax / Bcl- -myc mRNA were measured by semi-quantitative RT-PCR in HL-7702 / HBx and control group, respectively. RESULTS: RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. PCR analysis, Bax and c-myc mRNA levels in HL-7702 / HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702 / HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702 / H BX cells of transient and stable transfection were significantly lower than those in control. CONCLUSION: HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas / FasL, Bax / Bcl-2, and c-myc gene in a dose- dependent manner.