A mutant of hepatitis B virus X protein （HBx△127） enhances hepatoma cell migration via osteopontin involving 5-1ipoxygenase  

作　　者：Xuan ZHANG 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, Nankai University, Tianjin 300071, China [2]Department of Biochemistry, The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin 300071, China

出　　处：《Acta Pharmacologica Sinica》2010年第5期593-600,共8页中国药理学报（英文版）

基　　金：Acknowledgements This project was supported by grants from the National Basic Research Program of China （973 Program, No 2007CB914802, No 2007CB914804, No 2009CB521702） and the National Natura,1 Science Foundation （No 30670959）.

摘　　要：Aim： To explore a novel function of a mutant of the hepatitis B virus X protein （HBx△127） in the promotion of hepatoma cell migration. Methods： The effect of HBx△127 and wild type HBx on the migration ability of hepatoblastoma HepG2 cells were examined using wound healing assays in stable transfection systems. The full-length osteopontin（OPN） promoter sequence was cloned into the pGL3- Basic plasmid. The promoter activities of OPN in stably HBx△127-transfected hepatoblastoma HepG2 （HepG2-X△127） and hepatocellular carcinoma H7402 （H7402-X△127） cells were determined using luciferase reporter gene assays. The mRNA expression levels of OPN were detected by RT-PCR. And the effect of MK886, a specific inhibitor of 5-1ipoxygenase （5-LOX）, on OPN promoter activity and mRNA expression in HepG2-X△127 and H7402-X△127 cells were examined using luciferase reporter gene assays and RT-PCR, respectively. Finally, the migration ability of HepG2-X△127 was observed after treatment with siRNA targeting OPN mRNA and HBx mRNA using wound healing assays. Results： HepG2-X△127 cells exhibited a greater capacity for wound repair compared to HepG2-X cells. The promoter activity and mRNA expression levels of OPN were also increased in HepG2-X△127 and H7402-X△127 cells. Moreover, MK886 abolished the HBx△127- mediated upregulation of OPN. Wound healing assays demonstrated that the migration ability of HepG2-X△127 cells can be suppressed by treatment with siRNA targeting OPN mRNA and siRNA targeting HBx mRNA. Conclusion： HBx△127 strongly promotes hepatoma cell migration via activation of OPN involving 5-LOX.Aim： To explore a novel function of a mutant of the hepatitis B virus X protein （HBx△127） in the promotion of hepatoma cell migration. Methods： The effect of HBx△127 and wild type HBx on the migration ability of hepatoblastoma HepG2 cells were examined using wound healing assays in stable transfection systems. The full-length osteopontin（OPN） promoter sequence was cloned into the pGL3- Basic plasmid. The promoter activities of OPN in stably HBx△127-transfected hepatoblastoma HepG2 （HepG2-X△127） and hepatocellular carcinoma H7402 （H7402-X△127） cells were determined using luciferase reporter gene assays. The mRNA expression levels of OPN were detected by RT-PCR. And the effect of MK886, a specific inhibitor of 5-1ipoxygenase （5-LOX）, on OPN promoter activity and mRNA expression in HepG2-X△127 and H7402-X△127 cells were examined using luciferase reporter gene assays and RT-PCR, respectively. Finally, the migration ability of HepG2-X△127 was observed after treatment with siRNA targeting OPN mRNA and HBx mRNA using wound healing assays. Results： HepG2-X△127 cells exhibited a greater capacity for wound repair compared to HepG2-X cells. The promoter activity and mRNA expression levels of OPN were also increased in HepG2-X△127 and H7402-X△127 cells. Moreover, MK886 abolished the HBx△127- mediated upregulation of OPN. Wound healing assays demonstrated that the migration ability of HepG2-X△127 cells can be suppressed by treatment with siRNA targeting OPN mRNA and siRNA targeting HBx mRNA. Conclusion： HBx△127 strongly promotes hepatoma cell migration via activation of OPN involving 5-LOX.

关 键 词：hepatitis B virus X protein mutant of HBx cell migration OSTEOPONTIN HEPATOMA 5-1ipoxygenase 

分 类 号：R[医药卫生]