融合蛋白FADDdel-GFP对死亡受体介导的胰岛细胞凋亡的影响

Effects of fusion protein FADDdel-GFP on islet cell apoptosis mediated by death receptor

目的研究融合蛋白FADDdel-GFP对死亡受体介导的细胞凋亡信号的生物学效应,以探讨通过基因修饰靶细胞对1型糖尿病的影响。方法用脂质体将融合基因pFADDdel-GFP导入胰岛细胞株NIT,通过细胞RT-PCR和荧光显微镜检测其表达,采用FACS检测抗-Fas抗体诱导的胰岛细胞株的细胞毒效应。结果转染重组子pFADDdel-GFP的NITf-g细胞可见绿色荧光蛋白表达;NITf-g细胞对抗-Fas诱导的细胞损伤率为10.16%,细胞内的Caspase-3的活性为12.33%,均明显低于对照组(P<0.05)。结论成功建立了稳定表达FADDdel-GFP的胰岛细胞株;FADDdel-GFP可有效抑制死亡受体介导的细胞内凋亡信号传导。

Objective：To study biological effects of fusion protein FADDdel-GFP on the cell apoptosis signal mediated by death receptor, wheraby to investigate the influence on type 1 diabetes by modifying the target ceils with gene. Methods：After transfecting the fusional gene pFADDdel-GFP into mammalian ceils NIT（ mouse insulinoma ceils） by lipofectamine, the expression of the fusion protein was detected by RT-PCR and fluorescent microscope. Cytotoxicity on the islet ceils induced by anti-Fas was detected with FACS. Results：GFP was expressed in NIT transfected with pFADDdel-GFP. The percentage of ceil damage and the Caspase-3 activities in NITfg cells were 10. 16% and 12. 33% separately, both of which were lower than those of control groups obviously（ P 〈 0. 05 ）. Conclusiou：Sueecssfully established the islet ceil strain steadily expressing FADDdel-GFP. FADDdel-GFP could effectively inhibit the transduction of intracellular apoptosis signal by death receptor.