lncRNA Tmevpg1对小鼠自噬和JAK-STAT信号通路关键信号分子表达水平的影响

Effects of lncRNA Tmevpg1 on the Expression of Mouse Autophagy and Key Signaling Molecules in JAK-STAT Signaling Pathways

该文主要研究lncRNA Tmevpg1对小鼠自噬和JAK-STAT信号通路关键信号分子等相关分子表达水平的影响。该研究利用生物信息学分析构建Tmevpg1、JAK-STAT信号通路及自噬互作调控网络;雷帕霉素(rapamycin, Rapa)和氯喹(chloroquine, CQ)构建小鼠自噬模型;通过细胞共培养技术构建Tmevpg1过表达与干扰细胞模型;qRT-PCR(quantitative real-time PCR)检测小鼠脾脏和细胞模型中Tmevpg1等相关分子的RNA水平, Western blot检测自噬相关蛋白、T-bet和JAKSTAT通路关键信号分子磷酸化蛋白表达水平。结果显示, Tmevpg1、IFN-γ、T-bet、STAT1和JAK1在自噬发生的一定时间点内显著上调(P<0.001);过表达Tmevpg1后ULK1表达上调(P<0.05), p62下调(P<0.05), IFN-γ、T-bet、JAK1以及STAT1表达未发生明显改变,而干扰Tmevpg1后IFN-γ、T-bet、JAK1以及STAT1表达减少(P<0.05);Western blot结果显示小鼠自噬模型构建成功, T-bet、p-STAT1和p-JAK1表达趋势与m RNA水平一致;过表达Tmevpg1可上调ULK1和LC3-Ⅱ,下调p62,同时干扰Tmevpg1后T-bet、p-JAK1和p-STAT1表达水平下降。该项研究结果表明, lncRNA Tmevpg1和JAKSTAT信号通路对细胞自噬发挥重要调控作用。

This study was performed to explore the effect of lncRNA Tmevpg1 on the expression levels of mouse autophagy and key signaling molecules in JAK-STAT signaling pathway.In this study,Tmevpg1,JAKSTAT signaling pathway and autophagy interaction regulatory network were constructed by bioinformatics analysis;The mouse autophagy model was constructed with rapamycin(Rapa)and chloroquine(CQ),and the cell model of Tmevpg1 overexpression and interference was constructed by cell co-culture technique.The relative expression of Tmevpg1 and other related molecules were detected by qRT-PCR in mouse spleen issues and cell model,and Western blot verified the expression of autophagy-associated proteins,T-bet and phosphorylated proteins in key signaling molecules of JAK-STAT pathways.The results showed that Tmevpg1,IFN-γ,T-bet,STAT1 and JAK1 were significantly up-regulated at a certain time point of autophagy(P<0.001).After overexpression of Tmevpg1,ULK1 expression was up-regulated(P<0.05),p62 was down-regulated(P<0.05),IFN-γ,T-bet,JAK1 and STAT1 expression did not change significantly,but IFN-γ,T-bet,JAK1 and STAT1 expression were down-regulated after interference with Tmevpg1(P<0.05).Western blot results showed that the autophagy model of mice was successfully constructed,and the expression trends of T-bet,p-STAT1 and p-JAK1 were consistent with the level of mRNA.Overexpression of Tmevpg1 can up-regulate ULK1 and LC3-Ⅱand down-regulate p62.At the same time,the expression of T-bet,p-JAK1 and p-STAT1 decreased after interference with Tmevpg1.In conclusion,our research suggest that lncRNA Tmevpg1 and the JAK-STAT signaling pathway may play critical regulatory roles in autophagy.