人乳头瘤病毒16型E7基因靶向调控cGAS-STING信号通路抑制宫颈癌细胞免疫功能的研究

Human papillomavirus type 16 E7 gene targeted cGAS-STING signalingpathway to suppress immune function in cervical cancer cells

目的:探讨HPV16 E7靶向调控cGAS-STING信号通路对宫颈癌细胞免疫功能的影响。方法:体外构建HPV16 E7过表达和干扰质粒,转染至SiHa细胞,qPCR检测HPV16 E7 mRNA表达水平,流式细胞术检测CD95、MICA/B、CD73、CD39、CTLA-4和CD25表达。在此基础上分别转染cGAS-siRNA和STING-siRNA至SiHa细胞,CCK8检测细胞活力,流式细胞术检测细胞凋亡。qPCR和Western blot分别检测cGAS、STING、IFN-Ⅰ、HPV16 E7的mRNA及蛋白表达。流式细胞术检测细胞CD95、MICA/B、CD73、CD39、CTLA-4和CD25表达。结果:STING-siRNA+HPV16 E7过表达组CD95和MICA/B水平降低(P<0.05),CD73、CD39、CTLA-4和CD25水平升高(P<0.05)。cGAS、STING和IFN-Ⅰ蛋白表达水平降低(P<0.05)。STINGsiRNA+HPV16 E7干扰组结果与之相反。结论:HPV16 E7靶向调控cGAS-STING信号通路进而抑制宫颈癌细胞免疫功能。

Objective:To explore the effect of HPV16 E7 targeted cGAS-STING signaling pathway on immune function of cer-vical cancer cells.Methods:HPV16 E7 overexpression and interference plasmids were constructed in vitro and transfected into SiHa cells.mRNA expression level of HPV16 E7 was detected by qPCR,and expressions of CD95,MICA/B,CD73,CD39,CTLA-4 and CD25 were detected by flow cytometry.On this basis,cGAS-siRNA and STING-siRNA were transfected into SiHa cells respectively.Cell viability was detected by CCK8,and cell apoptosis was detected by flow cytometry.mRNA and protein expressions of cGAS,STING,IFN-Ⅰand HPV16 E7 were detected by qPCR and Western blot respectively.Expressions of CD95,MICA/B,CD73,CD39,CTLA-4 and CD25 were detected by flow cytometry.Results:Levels of CD95 and MICA/B were decreased in STING-siRNA+HPV16 E7 overexpression group(P<0.05),while levels of CD73,CD39,CTLA-4 and CD25 were increased(P<0.05).Protein expression levels of cGAS,STING and IFN-Ⅰwere decreased(P<0.05).Results of STING-siRNA+HPV16 E7 interference group were opposite.Conclusion:HPV16 E7 inhibits the immune function of cervical cancer cells by targeting cGAS-STING signaling pathway.