Screening of genes differentially expressed in HepG2 cells transfected with HCV NS3TP6 by microarray assay

    SHAO Qing, CHENG Jun, WANG Lin, ZHANG Jian, LU Chengª²Zhe

    1Gene Therapy Research Center, Institute of Infectious Diseases, PLA 302 Hospital, Beijing 100039, 2Department of CT, PLA 206 Hospital, Tonghua, Jilin 134001, China

    ¡¾Abstract¡¿ AIM: To study the difference in gene expression in human hepatoblastoma cell line HepG2 cells transfected with HCV NS3TP6ª²expressing plasmid and to further elucidate its molecular biological mechanism. METHODS: Sequence specific primers were designed and synthesized and the NS3TP6ª²coding DNA fragment was amplified with polymerase chain reaction (PCR) technique using pBRTM3011 plasmid containing the full length of HCVª²H cDNA as the template. The expression vector of pcDNA3.1(-)ª²NS3TP6 was constructed by routine molecular biological methods. cDNA microarray technology was employed to detect the mRNA from the HepG2 cells transfected respectively with pcDNA3.1(-)ª²NS3TP6 and pcDNA3.1(-) using lipofectamine. RESULTS: The expression vector was constructed and confirmed by restriction enzyme digestion and DNA sequencing analysis. The expression of NS3TP6 protein was confirmed by Western blot hybridization with single chain variable region (scFv) antibody. High quality mRNA and cDNA were prepared and successful microarray screening was conducted. The scanning results indicated that among the 1152 genes which were obtained from gene expression profile analysis, 45 were different, of which 7 genes were upª²regulated and 38 genes were downª²regulated in NS3TP6ª²expressing HepG2 cells. CONCLUSION: cDNA microarray technology is successfully used to screen the genes differentially expressed in NS3TP6ª²expressing HepG2 cells, which brings some new clues for the study of the molecular mechanism involved in the HCV infection and hepatocellular carcinoma development induced by HCV NS3TP6 protein. ......