The effect of HBx protein on gene transcription in Huh7 cells / 肿瘤

ABSTRACT

Objective: To study the effect of HBx on the gene expression profile in the hepatocellular carcinoma cell line Huh7 and to provide the molecular evidence for exploring the role of HBx in the tumorigenesis of hepatoma. Methods: HBx gene was transfected into Huh7 cells carried by pcDNA3. 1b plasmid which contained the entire open reading frame (ORF) of HBx gene. The empty vector of pcDNA3. 1b was used as a control. The HBx expression in transfected cells was confirmed by Western blotting as compared with those transfected with empty vectors. And then a microarray analysis was performed to detect the gene expression profile in Huh-7 cells by Human Genome U133 Plus 2.0 Array of Affymetrix-GeneChip. Six differentiated expressed genes (3 up-regulated and 3 down-regulated) were selected randomly and verified with RFQ-PCR. Results: The microarray analysis revealed that 88 genes were significantly up-regulated and 111 genes were down-regulated in Huh7 cells after transfection with HBx gene as compared with those transfected with empty vector. The functions of these differentiated genes were related with many aspects. Conclusion: HBx affected hepatocellular carcinoma cell HuH-7 bi-directionally. It not only had transcription-activating effects but also had transcription-suppressing effects. These data had important value for understanding the role of HBx in tumorigenesis and development of hepatoma.