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World J Gastroenterol. Oct 7, 2006; 12(37): 6046-6049
Published online Oct 7, 2006. doi: 10.3748/wjg.v12.i37.6046
Inhibition of hepatitis B virus expression and replication by RNA interference in HepG2.2.15
Zhong-Fu Zhao, Hui Yang, De-Wu Han, Long-Feng Zhao, Guo-Ying Zhang, Yun Zhang, Ming-She Liu
Zhong-Fu Zhao, Guo-Ying Zhang, Yun Zhang, Ming-She Liu, Institute of Hepatology, Changzhi Medical College, Changzhi 046000, Shanxi Province, China
Hui Yang, De-Wu Han, Long-Feng Zhao, Institute of Hepatology, Shanxi Medical University, Taiyuan 032000, Shanxi Province, China
Supported by Natural Science Foundation of Shanxi Province, China, No.20051114
Correspondence to: Dr. Zhong-Fu Zhao, Institute of Hepatology, Changzhi Medical College, Changzhi 046000, Shanxi Province, China. zhaozf_1226@163.com
Telephone: +86-355-3012335
Received: May 25, 2006
Revised: June 3, 2006
Accepted: June 16, 2006
Published online: October 7, 2006
Abstract

AIM: To observe the inhibition of hepatitis B virus replication and expression by transfecting vector-based small interference RNA (siRNA) pGenesil-HBV X targeting HBV X gene region into HepG2.2.15 cells.

METHODS: pGenesil-HBV X was constructed and transfected into HepG2.2.15 cells via lipofection. HBV antigen secretion was determined 24, 48, and 72 h after transfection by time-resolved immunofluorometric assays (TRFIA). HBV replication was examined by fluorescence quantitative PCR, and the expression of cytoplasmic viral proteins was determined by immunohistochemistry.

RESULTS: The secretion of HBsAg and HBeAg into the supernatant was found to be inhibited by 28.5% and 32.2% (P < 0.01), and by 38.67% (P < 0.05) and 42.86% (P < 0.01) at 48 h and 72 h after pGenesil-HBV X transfection, respectively. Immunohistochemical staining for cytoplasmic HBsAg showed a similar decline in HepG2.2.15 cells 48 h after transfection. The number of HBV genomes within culture supernatants was also significantly decreased 48 h and 72 h post-transfection as quantified by fluorescence PCR (P < 0.05).

CONCLUSION: In HepG2.2.15 cells, HBV replication and expression is inhibited by vector-based siRNA pGenesil-HBV X targeting the HBV X coding region.

Keywords: Hepatitis B virus, RNA interference, Plasmid vector, HepG2.2.15