FoxO1基因对Jurkat细胞FoxO1-KLF2-S1P1通路的调节作用  

Effects of FoxO1 on FoxO1-KLF2-S1P1 pathway in Jurkat cells

作　　者：闫飞 刘萍萍 张键 赵瑞 倪文鹏 李倩如 杜英 YAN Fei;LIU Ping-Ping;ZHANG Jian;ZHAO Rui;NI Wen-Peng;LI Qian-Ru;DU Ying(Department of Immunology,College of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450001,China)

机构地区：[1]郑州大学基础医学院免疫学系,郑州450001

出　　处：《中国免疫学杂志》2018年第3期335-339,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(81471545)

摘　　要：目的:通过构建FoxO1表达和干扰慢病毒载体,建立细胞内FoxO1-KLF2-S1P1信号通路调控研究模型,观察FoxO1过表达、干扰表达在Jurkat细胞内对其下游分子表达及功能的影响。方法:构建FoxO1表达和干扰表达慢病毒载体,分别感染Jurkat细胞,采用荧光定量PCR、Western blot和流式细胞术检测S1P1、CD62L、CCR7、CD69 mRNA水平和蛋白分子的表达。结果:FoxO1过表达组于感染后120 h FoxO1、KLF2、S1P1和CD62L mRNA水平显著增高(P<0.05),FoxO1、FoxO1-p和KLF2胞浆蛋白水平增高,S1P1^+细胞和CD62L^+细胞比率增高(P<0.05),CCR7^+细胞和CD69^+细胞未见显著改变(P>0.05)。FoxO1干扰组于转染后120 h FoxO1、KLF2、S1P1和CD62L mRNA水平降低(P<0.05),FoxO1、FoxO1-p和KLF2胞浆蛋白水平低于对照组,S1P1^+细胞百分比增多(P<0.05),但S1P1^+细胞和CD62L^+细胞在72 h时减少(P<0.05)。结论:FoxO1表达和干扰慢病毒载体转染Jurkat细胞并调节KLF2、S1P1和CD62L等分子的表达,为开展细胞内FoxO1-KLF2-S1P1信号通路调控和细胞相关功能的研究打下了基础。To observe the contribution of FoxO1 on its downstream molecules expression and function in Jurkat cells,Jurkat cells were infected with FoxO1 expression lentivirus and interference lentivirus to establish FoxO1-KLF2-S1P1 signaling pathways mod-el.After infection of 120 h,FoxO1,KLF2,S1P1 and CD62L mRNA levels and the protein expression of FoxO1,FoxO1-p and KLF2 were increased(P<0.05)in FoxO1-overexpression group.Converse results(P<0.05)were observed in the interference group.The proportions of S1P1+cells were increased in both groups.It was notably that S1P1+cells were decreased(P<0.05)in interference group after infection of 72 h.The proportion of CD62L+cells was increased(P<0.05)in overexpression group,it was decreased(P<0.05)in the interference group.This vitro experiments showed S1P1 and CD62L could be regulated by FoxO1 lentivirus,and provided an experimental basis for research about intracellular FoxO1-KLF2-S1P1 signaling pathways and cellular functions.

关 键 词：FOXO1 表达载体 干扰载体 S1P1 CD62L 

分 类 号：R392.11[医药卫生—免疫学]