Viral Hepatitis
Copyright ©The Author(s) 2004. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 1, 2004; 10(13): 1898-1901
Published online Jul 1, 2004. doi: 10.3748/wjg.v10.i13.1898
Effect of vector-expressed siRNA on HBV replication in hepatoblastoma cells
Jun Liu, Ying Guo, Cai-Fang Xue, Ying-Hui Li, Yu-Xiao Huang, Jin Ding, Wei-Dong Gong, Ya Zhao
Jun Liu, Cai-Fang Xue, Ying-Hui Li, Yu-Xiao Huang, Jin Ding, Wei-Dong Gong, Ya Zhao, Department of Etiology, Fourth Military Medical University, Xi’an 710033, Shaanxi Province, China
Ying Guo, Department of Obstetrics and Gynecology, Second Hospital of Xi’an Jiaotong University, Xi’an 710001, Shaanxi Province, China
Author contributions: All authors contributed equally to the work.
Supported by National Natural Science Foundation of China, No. 30100157; Medical Research Fund of Chinese PLA, No. 01MA184; and Innovation Project of FMMU, No. CX99005
Correspondence to: Dr. Jun Liu, Department of Etiology, Fourth Military Medical University, Xi’an 710033, Shaanxi Province, China. etiology@fmmu.edu.cn
Telephone: +86-29-83374536 Fax: +86-29-83374594
Received: December 12, 2003
Revised: January 8, 2004
Accepted: January 15, 2004
Published online: July 1, 2004
Abstract

AIM: To study the effect of siRNA expressed from DNA vector on HBV replication.

METHODS: Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.2.15 cells by using liposome-mediated transfection. 2.2.15 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed siRNA against green fluorescent protein and mock-transfected 2.2.15 cells were used as controls. Concentration of HBsAg in the supernatant of the transfected cells was measured by using solid-phase radioimmunoassay.

RESULTS: A mammalian siRNA expression vector pUC18U6 was constructed successfully. Compared with controls, pUC18U6HBVsir which expressed siRNA against HBV decreased concentration of HBsAg significantly by 44% (P < 0.05).

CONCLUSION: HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.

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