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Screening and identification of genes transactivated by hepatitis B virus X protein by microarray assay
Jun Cheng, Yan Liu, Yuan Hong, Jian-Jun Wang, Qian Yang
Jun Cheng, Yan Liu, Yuan Hong, Jian-Jun Wang, Qian Yang, Gene Therapy Research Center, Institute of Infectious Diseases, The 302 Hospital of PLA, Beijing, China
Supported by: Grants from National Natural Science Foundation No. C03011402, No. C30070689, and Returned Scholarship of General Logistics Department of PLA.
Correspondence to: Dr. Jun Cheng, Professor, Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, 100 Xisihuanzhong Road, Beijing 100039, China. cj@genetherapy.com.cn
Received: January 11, 2003 Revised: February 20, 2003 Accepted: March 21, 2003 Published online: July 15, 2003
AIM
Hepatitis B virus X protein (HBxAg) is a strong transactivator implicating in the transformation of normal hepatocyte and carcinogenesis. To understand the molecular mechanism of HBxAg in the up- and down-regulated genes of hepatocyte, we conducted microarray assay.
METHODS
The coding sequence of HBxAg was amplified by polymerase chain reaction (PCR) by using pCP10 containing the full length HBV genome DNA as the template. The expression of HBxAg in the transfected HepG2 cells was confirmed by Western blot methods. Total mRNA was isolated from the transfected HepG2 cells with pcDNA3.1(-) and pcDNA3-HBxAg, respectively. cDNA was prepared by reverse transcription. Microarray was conducted for screening of up- and down-regulated genes of both HepG2 cells.
RESULTS
The expressive vector of pcDNA3-HBxAg has been constructed and confirmed by restriction enzyme digestion and DNA sequencing. The expression of HBxAg has been confirmed by Western blot analysis. After screening with DNA microarray, we found 16 genes have been up-regulated, and 58 genes have been down-regulated.
CONCLUSION
The HBxAg is a very strong transactivator. Microarray is an important choice for the analysis of target genes of HBxAg transactivation.
Key Words: N/A
Citation: Cheng J, Liu Y, Hong Y, Wang JJ, Yang Q. Screening and identification of genes transactivated by hepatitis B virus X protein by microarray assay. Shijie Huaren Xiaohua Zazhi 2003; 11(7): 920-924
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