Gene expression profiles of human liver cells mediated by hepatitis B virus X protein  

作　　者：Wei-ying ZHANG Fu-qing XU Chang-liang SHAN Rong XIANG Li-hong YE Xiao-dong ZHANG 

机构地区：[1]Department of Cancer Research, Key Laboratory of Molecular Microbiology and Technology of Ministry of Education, Institute for Molecular Biology, College of Life Sciences [2]Department of Biochemistry, College of Life Sciences [3]School of Medicine, Nankai University, Tianjin 300071, China

出　　处：《Acta Pharmacologica Sinica》2009年第4期424-434,共11页中国药理学报（英文版）

基　　金：This project was supported by grants from the National Basic Research Program of China （973 Program, No 2007CB914802, No 2007CB914804, No 2009CB521702）, Chinese State Key Projects for High-Tech 863 Program （2006AA02A247）,and the National Natural Science Foundation （No 30670959）.

摘　　要：Aim： To demonstrate the gene expression profiles mediated by hepatitis B virus X protein （HBx）, we characterized the molecular features of pathogenesis associated with HBx in a human liver cell model. Methods： We examined gene expression profiles in L-O2-X cells, an engineered L-O2 cell line that constitutively expresses HBx, relative to L-O2 cells using an Agilent 22 K human 70-mer oligonucleotide microarray representing more than 21,329 unique, well-characterized Homo sapiens genes. Western blot analysis and RNA interference （RNAi） targeting HBx mRNA validated the overexpression of proliferating cell nuclear antigen （PCNA） and Bcl-2 in L-O2-X cells. Meanwhile, the BrdU incorporation assay was used to test cell proliferation mediated by upregulated cyclooxygenase-2 （COX-2）. Results： The microarray showed that the expression levels of 152 genes were remarkably altered; 82 of the genes were upregulated and 70 genes were downregulated in L-O2-X cells. The altered genes were associated with signal transduction pathways, cell cycle, metastasis, transcriptional regulation, immune response, metabolism, and other processes. PCNA and Bcl-2 were upregnlated in L-O2-X cells. Furthermore, we found that COX-2 upregulation in L-O2-X cells enhanced prolif- eration using the BrdU incorporation assay, whereas indomethacin （an inhibitor of COX-2） abolished the promotion. Conclusion Our findings provide new evidence that HBx is able to regulate many genes that may be involved in the carcinogenesis. These regulated genes mediated by HBx may serve as molecular targets for the prevention and treatment of hepatocellular carcinoma.Aim： To demonstrate the gene expression profiles mediated by hepatitis B virus X protein （HBx）, we characterized the molecular features of pathogenesis associated with HBx in a human liver cell model. Methods： We examined gene expression profiles in L-O2-X cells, an engineered L-O2 cell line that constitutively expresses HBx, relative to L-O2 cells using an Agilent 22 K human 70-mer oligonucleotide microarray representing more than 21,329 unique, well-characterized Homo sapiens genes. Western blot analysis and RNA interference （RNAi） targeting HBx mRNA validated the overexpression of proliferating cell nuclear antigen （PCNA） and Bcl-2 in L-O2-X cells. Meanwhile, the BrdU incorporation assay was used to test cell proliferation mediated by upregulated cyclooxygenase-2 （COX-2）. Results： The microarray showed that the expression levels of 152 genes were remarkably altered; 82 of the genes were upregulated and 70 genes were downregulated in L-O2-X cells. The altered genes were associated with signal transduction pathways, cell cycle, metastasis, transcriptional regulation, immune response, metabolism, and other processes. PCNA and Bcl-2 were upregnlated in L-O2-X cells. Furthermore, we found that COX-2 upregulation in L-O2-X cells enhanced prolif- eration using the BrdU incorporation assay, whereas indomethacin （an inhibitor of COX-2） abolished the promotion. Conclusion Our findings provide new evidence that HBx is able to regulate many genes that may be involved in the carcinogenesis. These regulated genes mediated by HBx may serve as molecular targets for the prevention and treatment of hepatocellular carcinoma.

关 键 词：hepatitis B virus HBX MICROARRAY COX-2 PROLIFERATION 

分 类 号：R373.21[医药卫生—病原生物学] R734.2[医药卫生—基础医学]