融合基因FADDdel-GFP修饰在胰岛细胞自杀效应中的作用

Roles of fusion gene pFADDdel-GFP modification in self-death of islet cell

目的研究FADDdel-GFP在Fas/FasL介导的胰岛细胞自杀效应中的作用。方法采用DNA转染技术将重组体pFADDdel-GFP导入哺乳动物细胞NIT(鼠胰岛细胞瘤细胞);采用FACS检测细胞因子诱导后NIT细胞的Fas和FasL表达情况及细胞自杀效应;采用active caspase3检测试剂盒检测细胞因子作用前后NIT细胞的caspase-3活性变化。结果NIT细胞表面无Fas和FasL表达,细胞因子可促其表达增加;细胞因子作用后,FADDdel-GFP修饰的NIT细胞的细胞损伤百分率和细胞内caspase3活性明显低于对照组。结论FADDdel-GFP修饰的NIT细胞具有一定的抵抗Fas/FasL途径介导细胞自杀效应的能力。

Objective To study the role of FADDdel-GFP in Fas/FasL-mediated self-death of islet cell. Methods The recombinant vector pFADDdel-GFP was transfected into mammalian cells NIT. The levels of Fas and FasL expression on NIT cells and the cell self-death effect induced by cytokines were detected by FAGS. Changes of the caspase-3 activity in NIT cell with or without cytokine induction were detected by active caspase3 kit. Results There were no Fas attd FasL expressions on NIT cells. But cytokine could promote the expressions of Fas and FasL. The cell damage percentage and caspase3 activity of NIT modified with FADDdel-GFP were lower than those of control groups. FADDdel-GFP-modified NIT cells have the capability of resistance to cell self-death mediated by Fas/FasL pathway.