Rapid Communication
Copyright ©2007 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Apr 28, 2007; 13(16): 2324-2327
Published online Apr 28, 2007. doi: 10.3748/wjg.v13.i16.2324
Combination of small interfering RNA and lamivudine on inhibition of human B virus replication in HepG2.2.15 cells
Gui-Qiu Li, Wei-Zhen Xu, Jing-Xia Wang, Wen-Wei Deng, Di Li, Hong-Xi Gu
Gui-Qiu Li, Wei-Zhen Xu, Di Li, Hong-Xi Gu, Department of Microbiology, Harbin Medical University, Harbin 150081, Heilongjiang Province, China
Jing-Xia Wang, Department of Histology and Embryology, Jiamusi Medical College, Jiamusi University, Jiamusi 154002, Heilongjiang Province, China
Wen-Wei Deng, Department of Anesthesiology, Second Affiliated Hospital, Jiamusi University, Jiamusi 154002, Heilongjiang Province, China
Author contributions: All authors contributed equally to the work.
Supported by PhD Foundation of Education Ministry, China, No. 2005006; Youth Foundation of Heilongjiang Province, No. QC060061; and Foundation of Health Hall, Heilongjiang Province, No. 2005-009
Correspondence to: Hong-Xi Gu, Department of Microbiology, Harbin Medical University, Harbin 150081, Heilongjiang Province, China. hxgu2432@163.com
Telephone: +86-451-86685122
Received: January 18, 2007
Revised: February 3, 2007
Accepted: March 28, 2007
Published online: April 28, 2007
Abstract

AIM: To observe the inhibition of hepatitis B virus (HBV) replication and expression by combination of siRNA and lamivudine in HepG2.2.15 cells.

METHODS: Recombinant plasmid psil-HBV was constructed and transfected into HepG2.2.15 cells. The transfected cells were cultured in lamivudine-containing medium (0.05 μmol/L) and harvested at 48, 72 and 96 h. The concentration of HBeAg and HBsAg was determined using ELISA. HBV DNA replication was examined by real-time PCR and the level of HBV mRNA was measured by RT-PCR.

RESULTS: In HepG2.2.15 cells treated with combination of siRNA and lamivudine, the secretion of HBeAg and HBsAg into the supernatant was found to be inhibited by 91.80% and 82.40% (2.89 ± 0.48 vs 11.73 ± 0.38, P < 0.05; 4.59 ± 0.57 vs 16.25 ± 0.48, P < 0.05) at 96 h, respectively; the number of HBV DNA copies within culture medium was also significantly decreased at 96 h (1.04 ± 0.26 vs 8.35 ± 0.33, P < 0.05). Moreover, mRNA concentration in HepG2.2.15 cells treated with combination of siRNA and lamivudine was obviously lower compared to those treated either with siRNA or lamivudine (19.44 ± 0.17 vs 33.27 ± 0.21 or 79.9 ± 0.13, P < 0.05).

CONCLUSION: Combination of siRNA and lamivudine is more effective in inhibiting HBV replication as compared to the single use of siRNA or lamivudine in HepG2.2.15 cells.

Keywords: Hepatitis B virus; RNA interference; siRNA with lamivudine; HepG2.2.15 cell