pEGFP-N1-IL-17通过调控Fas/FasL信号通路影响喉癌Hep-2细胞的凋亡

pEGFP-N1-IL-17 regulates Fas/FasL pathway and affects apoptosis of laryngeal carcinoma cells

目的:研究重组质粒白介素17(pEGFP-N1-IL-17)对喉癌细胞(Hep-2细胞)凋亡的影响,阐述pEGFP-N1-IL-17通过Fas/FasL信号通路抑制Hep-2细胞凋亡的机制。方法:设置未转染的Hep-2细胞为空白对照组,pEGFP-N1-NC转染的喉癌Hep-2细胞为阴性对照组,pEGFP-N1-IL-17转染的喉癌Hep-2细胞为实验组。qRT-PCR、Western blot实验用来检测IL-17在Hep-2细胞的表达水平,成功构建了IL-17过表达载体。用Western blot检测空白对照组、阴性对照组、实验组中Fas、FasL、Cleaved-Caspase3、Cleaved-Caspase8蛋白的表达情况。用流式细胞仪检测空白对照组、阴性对照组、实验组细胞的凋亡率。结果:IL-17过表达载体转染喉癌Hep-2细胞后,qRT-PCR检测IL-17表达水平上调数倍。与阴性对照组相比,空白对照组Fas、FasL、Cleaved-Caspase3、Cleaved-Caspase8蛋白表达无明显差异。与阴性对照组相比,实验组喉癌Hep-2细胞内Fas、FasL、Cleaved-Caspase3、Cleaved-Caspase8蛋白表达明显降低,差异有统计学意义(P<0.01)。空白对照组、阴性对照组的细胞凋亡率没有明显变化。与空白对照组相比,实验组的喉癌Hep-2细胞的凋亡率有明显降低。结论:IL-17可能通过Fas/FasL信号通路来抑制喉癌Hep-2细胞凋亡。

Objective To investigate the effect of pEGFP-N1-IL-17 on the apoptosis cells and to explain the mechanism by which pEGFP-N1-IL-17 inhibits the apoptosis of Hep-2 cells through the Fas/FasL signaling pathway.Methods The recombinant plasmid vector pEGFP-N1-IL-17 carrying IL-17 gene was constructed and transfected into Hep-2 cells,normal Hep-2 cells were used as the control group,and pEGFP-N1-NC was used as negative control group.pEGFP-N1-IL-17 was used as the experimental group.qRT-PCR and Western blot were used to detect the expression level of IL-17 in Hep-2 cells,and IL-17 overexpression vector was successfully constructed.The effect of pEGFP-N1-IL-17 on the apoptosis rate of Hep-2 cells was detected by flow cytometry.Western blot was used to detect the expression of Fas and FasL in Hep-2 cells transfected by pEGFP-N1-IL-17.Results After IL-17 overexpression vector was transfected into laryngeal carcinoma Hep-2 cells,the expression level of IL-17 was up-regulated by qRT-PCR.Annexin V-FITC/PI cell apoptosis assay results showed that the apoptosis rate of Hep-2 cells in experimental groups decreased obviously.The results of Western blot showed that the expressions of Fas,FasL,Cleaved-Caspase8 and Cleaved-Caspase3 in experinental group of Hep-2 cells were significantly decreased(P<0.01).Conclusion IL-17 may inhibit the apoptosis of Hep-2 cells through Fas/FasL signaling pathway.