基础医学与临床 ›› 2018, Vol. 38 ›› Issue (12): 1718-1723.

• 研究论文 • 上一篇    下一篇

乙型肝炎病毒反式激活因子XTP4促进肝癌细胞系迁移和侵袭

邓秀娟1,韩铭2,刘顺爱2,成军2,梁跃东3   

  1. 1. 贵州医科大学
    2. 首都医科大学附属北京地坛医院传染病研究所
    3. 贵阳医学院
  • 收稿日期:2017-11-24 修回日期:2018-02-01 出版日期:2018-12-05 发布日期:2018-11-23
  • 通讯作者: 梁跃东 E-mail:lyd302@163.com
  • 基金资助:
    北京市医管局“登峰”人才培养计划团队项目;重点医学专业发展计划项目

HBX protein trans-activiate gene XTP4 promotes the migration and invasion of hepatocellular carcinoma cell line

  • Received:2017-11-24 Revised:2018-02-01 Online:2018-12-05 Published:2018-11-23

摘要: 目的 探讨在肝癌细胞株HepG2中过表达和干扰XTP4表达后,对细胞迁移和侵袭能力的影响。方法 实验分对照组、过表达组和干扰组,将构建成功的XTP4质粒和化学合成的XTP4小干扰RNA(siRNA)瞬时转染入HepG2,培养48h后,实时荧光定量PCR(q RT-PCR)和蛋白印迹(western-blot)检测细胞内XTP4 mRNA和蛋白表达;Snail和NF-κB以及上皮间质转化EMT相关分子的表达。并通过细胞划痕愈合实验、Transwell迁移侵袭实验观察细胞迁移侵袭能力。结果 成功重培养并提取出XTP4质粒;过表达组XTP4的转录水平和翻译水平均明显高于对照组,迁移和侵袭能力明显增加,下游分子NF-κB、Snail和MMP-9表达均增加。干扰组XTP4的转录水平和翻译水平较对照组均明显降低,迁移和侵袭能力也明显降低,NF-κB、Snail和MMP-9表达减少。结论 乙型肝炎病毒反式激活因子XTP4可促进HepG2迁移和侵袭,与NF-κB、Snail和MMP-9调节相关,EMT相关分子E-cadherin、N-cadherin可能参与其中。

关键词: XTP4基因, 肝癌细胞, 迁移侵袭

Abstract: Objective To investigate the effect of over-expression of XTP4 and the interference of XTP4 on cell migration and invasion in HepG2. Methods The experiment was divided into control group, overexpression group and interference group. The constructed recombinant plasmid XTP4 and the chemically synthesized XTP4 small interfering RNA (siRNA) were transiently transfected into HepG2 cells and cultured for 48 hours. Real-time quantitative PCR (q RT-PCR) Western blotting was used to detect the expression of XTP4 mRNA and protein, Snail and NF-κB as well as EMT-related molecules in epithelial-mesenchymal transition. Through cell scratch healing experiment and Transwell migration and invasion assay, the cell migration and invasion ability was observed. Results The XTP4 plasmid was successfully re-cultured and extracted. The transcription and translation levels of XTP4 in overexpression group were significantly higher than those in control group. The migration and invasion ability of XTP4 were significantly increased, while the expressions of NF-κB, Snail and MMP-9 were also increased. The transcription level and translation level of XTP4 in the interference group were significantly lower than those in the control group, the ability of migration and invasion was also significantly decreased, and the expressions of NF-κB, Snail and MMP-9 were decreased. Conclusions Hepatitis B virus transactivator XTP4 may promote the migration and invasion of HepG2, which may be related to the regulation of NF-κB, Snail and MMP-9. EMT-related molecules E-cadherin and N-cadherin may be involved.

Key words: XTP4 gene, hepatoma cell, migration and invasion