Hepatitis B virus preS2 activates human acyl protein thioesterase 1 promoter / 重庆医学

ABSTRACT

Objective To investigate the trans‐regulative effect of hepatitis B virus (HBV) preS2 on the promoter of human acyl protein thioesterase 1 (APT1) gene .Methods The promoter sequence of human APT1 gene was identified applying the soft‐ware of bioinformatics .The APT1 promoter and HBV preS2 gene were amplified with PCR and cloned into pGL3 and pcDNA3 .1 (-) plasmids to construct the luciferase reporter gene plasmid of human APT1 gene promoter pGL3‐APT1 and the preS2 eukary‐otic expression plasmid pcDNA3 .1(-)‐preS2 ,respectively .The effect of the preS2 on the human APT1 gene promoter was exam‐ined by cotransfecting hepatocellular carcinoma cell HepG2 with pGL3‐APT1 and pcDNA3 .1(-)‐preS2 and measuring luciferase activities of the HepG2 cells .The statistical data were analyzed with independent‐samples t test .Results Both plasmids of pGL3‐APT1 and pcDNA3 .1(-)‐preS2 were confirmed by DNA sequencing to be accurately constructed as design .The luciferase activity of the pGL3‐APT1 was 1 .2 times (P<0 .01) that of the positive control plasmid pGL3‐Control .And the luciferase activity of the HepG2 cells cotransfected with pcDNA3 .1(-)‐preS2 and pGL3‐APT1 was 2 .6 times (P<0 .01) that of the HepG2 cells cotrans‐fected with the plasmid without preS2 gene pcDNA3 .1(-) and pGL3‐APT1 .Conclusion The human APT1 promoter cloned in the study has high promoter activity ;HBV preS2 activates human APT1 promoter .