Inhibition of human La protein by RNA interference downregulates hepatitis B virus mRNA in 2.2.15 cells.

ABSTRACT:

Aim

To investigate the role of human La protein in HBV mRNA expression.

Methods

Three human La protein (hLa) specific siRNA expression cassettes (SECs) containing U6+1 promoter were prepared via one-step overlapping extension PCR. After transfection with SECs into HepG2 cells, inhibition effects on hLa expression were analyzed by semi-quantitative RT-PCR and Western blotting. Then, effective SECs were screened out and transfected into 2.2.15 cells, a stable HBV-producing cell line. HBV surface antigen (HBsAg) and e antigen (HBeAg) secretions into culture media were detected by microparticle enzyme immunoassay (MEIA) and HBs and HBe mRNA levels were analyzed by semi-quantitative RT-PCR.

Results

SEC products containing U6+1 snRNA promoter, and 3 sites of hLa mRNA specif

SUBMITTER: Ni Q 

PROVIDER: S-EPMC4572331 | biostudies-literature | 2004 Jul

REPOSITORIES: biostudies-literature