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    鸭坦布苏病毒E蛋白及其结构域I、II、III对DEF细胞周期与凋亡的影响

    Effects of duck Tembutsu virus E protein and its domains of I, II, III on DEF cell cycle and apoptosis

      摘要
    • 摘要:
      目的  探究鸭坦布苏病毒(Duck Tambusu virus，DTMUV) E蛋白全长及其结构域I、II和III (DI、DII和DIII)对鸭胚成纤维细胞(Duck embryo fibroblast，DEF)的细胞周期与凋亡的影响。
      方法  本研究设计、合成DTMUV E蛋白全长及其DI、DII和DIII真核表达质粒，并转染至DEF，用流式细胞仪检测不同蛋白引起的DEF细胞凋亡和细胞周期的变化。
      结果  细胞凋亡结果显示：质粒转染细胞24 h后，E蛋白全长及DI、DII、DIII诱导的DEF早期凋亡率分别是16.4%、15.1%、14.0%和17.2%；质粒转染细胞36 h后，E蛋白全长及DI、DII、DIII诱导的DEF早期凋亡率分别是23.4%、18.5%、26.7%和29.4%。细胞周期检测结果显示：E蛋白全长及DI、DII、DIII的质粒转染细胞24、36 h 后，DNA合成期(S期)细胞比例都明显高于pEGFP-N1空载体转染组。质粒转染24 h后，E蛋白及DI、DII、DIII的S期细胞比例分别为5.43%、22.58%、12.75%和12.80%；质粒转染细胞36 h后，E蛋白及DI、DII、DIII的S期细胞比例分别为9.98%、11.44%、10.44%和11.00%。
      结论  DTMUV E蛋白全长及其3个结构域均能诱导早期细胞凋亡，引起细胞S期的停滞，但是各蛋白片段诱导细胞凋亡和引起细胞周期变化的能力存在一定差异。

       

      Abstract:
      Objective  To investigate the effects of the whole length of Duck Tambusu virus (DTMUV) E protein and its domains of I, II and III (DI, DII and DIII) on the cell cycle and apoptosis of duck embryo fibroblast (DEF) cells.
      Method  The full-length of DTMUV E protein, DI, DII and DIII eukaryotic expression plasmids were designed and synthesized, and transfected into DEF. The changes of DEF apoptosis and cell cycle caused by different proteins were detected by flow cytometry.
      Result  The results of apoptosis showed that after 24 h of plasmid transfection, the apoptosis rates of DEF induced by the full length, DI, DII and DIII were 16.4%, 15.1%, 14.0% and 17.2% respectively; after 36 h of plasmid transfection, the apoptosis rates of DEF induced by E protein full length, DI, DII and DIII were 23.4%, 18.5%, 26.7% and 29.4%. The results of cell cycle detection showed that the percentage of DNA synthesis phase (S-phase) cells transfected with E protein full length, DI, DII and DIII plasmid was obviously higher than those of cells transfected with pEGFP-N1 empty vector 24 h and 36 h later. After 24 h of plasmid transfection, the percentage of S-phase cells in E protein, DI, DII and DIII groups were 5.43%, 22.58%, 12.75% and 12.80%, respectively; After 36 h of plasmid transfection, the percentage of S-phase cells in E protein, DI, DII and DIII groups were 9.98%, 11.44%, 10.44% and 11.00%, respectively.
      Conclusion  The full length of DTMUV E protein and its three domains can induce early cell apoptosis and arrest the S-phase cells, but the ability of each protein fragment to induce cell apoptosis and change the cell cycle is different.

       

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