Liu Li-ping, Chen Xiao-ping, Zhang Wan-guang, Yang Sheng-li, Liang Hui-fang, Xu Tao, Ren Li, Zhang Wei
Hepatic Surgery Center, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, China.
Zhonghua Wai Ke Za Zhi. 2008 Jul 15;46(14):1092-6.
To study the function of nuclear factor-kappaB (NF-kappaB) signaling pathway in regulating vascular endothelial growth factor (VEGF) by hepatitis B virus X protein (HBx).
After the establishment of L02-HBx cell line with stable transfected HBx gene, NF-kappaB signaling pathway blocker PDTC was introduced to cut off its signal transduction. Double immunofluorescent staining and laser scanning confocal microscopy were applied to study the activation and deactivation of NF-kappaB signaling pathway. Real-time PCR and Western blot were used to observe the expression of VEGF gene before and after the HBx transfection, as well as the treatment with PDTC.
The NF-kappaB signaling pathway of L02-HBx cells was activated after transfection with HBx gene as compared to the control L02 cells without treatment. The mRNA and protein levels of VEGF in L02-HBx cells increased 4.07 +/- 0.31 and 4.34 +/- 0.64 times respectively. The difference was of statistical significance (P < 0.05) in comparison with the control cells. The mRNA levels of VEGF decreased to 2.33 +/- 0.22 and 1.86 +/- 0.18(P < 0.05) and at the same time the expression of VEGF also reduced to 2.52 +/- 0.29 and 2.17 +/- 0.34 (P < 0.05), after treatment with 25.0 micromol/L and 50.0 micromol/L PDTC for 24 h respectively when the NF-kappaB signaling pathway was blocked. There was no significant difference in VEGF mRNA and protein levels when treated with 12.5 micromol/L PDTC for 24 h.
NF-kappaB signaling pathway maybe one of the routes through which HBx up-regulate the expression of VEGF to promote angiogenesis in hepatocellular carcinoma.
研究乙型肝炎病毒X蛋白（HBx）通过核因子-κB（NF-κB）信号通路调控血管内皮生长因子（VEGF）的作用。
构建稳定转染HBx基因的L02-HBx细胞系，加入NF-κB信号通路阻断剂PDTC切断其信号转导。采用双免疫荧光染色及激光扫描共聚焦显微镜观察NF-κB信号通路的激活与失活情况。运用实时荧光定量PCR和蛋白质免疫印迹法观察转染HBx基因前后以及PDTC处理后VEGF基因的表达情况。
与未处理的对照L02细胞相比，转染HBx基因后L02-HBx细胞的NF-κB信号通路被激活。L02-HBx细胞中VEGF的mRNA和蛋白水平分别升高了4.07±0.31倍和4.34±0.64倍。与对照细胞相比，差异具有统计学意义（P<0.05）。分别用25.0 μmol/L和50.0 μmol/L的PDTC处理24 h阻断NF-κB信号通路后，VEGF的mRNA水平分别降至2.33±0.22和1.86±0.18（P<0.05），同时VEGF的表达也分别降至2.52±0.29和2.17±0.34（P<0.05）。用12.5 μmol/L的PDTC处理24 h后，VEGF的mRNA和蛋白水平无显著差异。
NF-κB信号通路可能是HBx上调VEGF表达以促进肝癌血管生成的途径之一。
