Inhibition of HBV transcription in HepG2.2.15 cells by artificial Zinc finger protein designed specifically to target HBX gene promoter

Abstract:

Objective：To observe the inhibition of hepatitis B virus（HBV） transcription in vitro by Zinc finger protein（ZFP） which was designed specifically to target X gene promoter of HBV（HBX）. Methods：Recombinant plasmid pcDNA3.1-ZFP was transfected into HepG2.2.15 cells. Expression of HBX protein was detected by Western blot at 24 h after being transfected. Hepatitis B e antigen（HBeAg） and HBV DNA levels in supernatant of HepG2.2.15cells at 24 h after being transfected with pcDNA3.1-ZFP were detected by ELISA and real-time PCR. HBV mRNA was tested by RT-PCR. Results：ZFP could inhibit the expression of HBX in HepG2.2.15 cells. In the presence of ZFP，HBV DNA level and HBeAg was considerably reduced by 51.7%（t=23.079，P=0.000，９５％CI=44.98%-58.52%） and 33.8%（t=3.887，P=0.003，９５％CI=12.12%-55.48%） respectively，while HBV mRNA was sharply decreased（t=3.616，P=0.022）. Conclusion：ZFP protein designed to target X gene promoter of HBV can inhibit the transcription of HBV in HepG2.2.15 cells effectively.