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.
Acta Pharmacol Sin. 2010 May;31(5):593-600. doi: 10.1038/aps.2010.36. Epub 2010 Apr 5.
To explore a novel function of a mutant of the hepatitis B virus X protein (HBx Delta 127) in the promotion of hepatoma cell migration.
The effect of HBx Delta 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 Delta 127-transfected hepatoblastoma HepG2 (HepG2-X Delta 127) and hepatocellular carcinoma H7402 (H7402-X Delta 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-lipoxygenase (5-LOX), on OPN promoter activity and mRNA expression in HepG2-X Delta 127 and H7402-X Delta 127 cells were examined using luciferase reporter gene assays and RT-PCR, respectively. Finally, the migration ability of HepG2-X Delta 127 was observed after treatment with siRNA targeting OPN mRNA and HBx mRNA using wound healing assays.
HepG2-X Delta 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 Delta 127 and H7402-X Delta 127 cells. Moreover, MK886 abolished the HBx Delta 127-mediated upregulation of OPN. Wound healing assays demonstrated that the migration ability of HepG2-X Delta 127 cells can be suppressed by treatment with siRNA targeting OPN mRNA and siRNA targeting HBx mRNA.
HBx Delta 127 strongly promotes hepatoma cell migration via activation of OPN involving 5-LOX.
探索乙型肝炎病毒 X 蛋白（HBxΔ127）突变体在促进肝癌细胞迁移中的新功能。
采用稳定转染系统中的划痕愈合实验，研究 HBxΔ127 和野生型 HBx 对肝癌 HepG2 细胞迁移能力的影响。将全长骨桥蛋白（OPN）启动子序列克隆到 pGL3-Basic 质粒中。通过荧光素酶报告基因检测稳定转染 HBxΔ127 的肝癌 HepG2（HepG2-XΔ127）和肝癌 H7402（H7402-XΔ127）细胞中 OPN 的启动子活性。通过 RT-PCR 检测 OPN 的 mRNA 表达水平。通过荧光素酶报告基因检测和 RT-PCR 分别检测特异性 5-脂氧合酶（5-LOX）抑制剂 MK886 对 HepG2-XΔ127 和 H7402-XΔ127 细胞中 OPN 启动子活性和 mRNA 表达的影响。最后，通过划痕愈合实验观察靶向 OPN mRNA 和 HBx mRNA 的 siRNA 处理后 HepG2-XΔ127 的迁移能力。
与 HepG2-X 细胞相比，HepG2-XΔ127 细胞具有更强的伤口修复能力。HepG2-XΔ127 和 H7402-XΔ127 细胞中 OPN 的启动子活性和 mRNA 表达水平也增加。此外，MK886 消除了 HBxΔ127 介导的 OPN 上调。划痕愈合实验表明，靶向 OPN mRNA 和靶向 HBx mRNA 的 siRNA 处理可抑制 HepG2-XΔ127 细胞的迁移能力。
HBxΔ127 通过激活涉及 5-LOX 的 OPN 强烈促进肝癌细胞迁移。
