Inhibition of hepatitis B virus replication in 2.2.15 cells by expressed shRNA

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Abstract
Summary. Hepatitis B virus (HBV) infection is a worldwide health problem. To determine whether RNA interference (RNAi) could inhibit ongoing HBV replication in 2.2.15 cells, we constructed shRNA‐producing vector pU6P based on the mouse U6 RNA promoter and cloned 12 targeted sequences against HBV into the vector, resulting in a series of pU6‐siHBV vectors. The recombinant vectors were transfected into 2.2.15 cells, HBsAg and HBeAg in cultured media were assayed using enzyme‐linked immunosorbent assay at various days after transfection. The amount of HBV DNA in the culture medium was quantitated by real‐time polymerase chain reaction. HBsAg and HBeAg expression were inhibited by 72.8 ± 5.4% (P = 0.00003) and 55.8 ± 6.2% (P = 0.000026), respectively, 4 days after transfection with pU6‐siHBV5. The greatest inhibition of HBV DNA was decreased by approximately 1.9‐fold (P = 0.013) on day 6 post transfection with pU6‐siHBV11 compared with that of empty vector. No change was found for HBV protein expression and DNA replication on pU6‐siGFP (negative control) transfected cells. Our data demonstrate that the transfection of HBV‐targeted shRNA‐producing vector in 2.2.15 cells could inhibit the HBV protein expression and HBV DNA replication specifically. RNAi may be considered as a potential antiviral approach for human HBV infection.