临床肝胆病杂志2025，Vol.41Issue(12)：2572-2580,9.DOI:10.12449/JCH251219
环磷酸鸟苷-腺苷合成酶-干扰素基因刺激因子(cGAS-STING)信号通路激活对γδT细胞杀伤肝癌细胞效应的调控作用
Activation of the cyclic guanosine monophosphate-adenosine monophosphate adenosine synthetase-stimulator of interferon genes signaling pathway regulates the cytotoxicity of γδT cells against hepatoma cells
摘要
Abstract
Objective To investigate the regulatory effect of the cyclic guanosine monophosphate-adenosine monophosphate adenosine synthetase(cGAS)-stimulator of interferon genes(STING)signaling pathway on the cytotoxicity of γδT cells against hepatocellular carcinoma(HCC)through in vitro experiments,and to provide new ideas for improving the efficacy of adoptive immunotherapy based on γδT cells.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and γδT cells were multiplied,and their purity was measured.Mature γδT cells were divided into γδT group,γδT-G10 group,and γδT-H-151 group.After 24 hours of in vitro stimulation,Western blot was used to measure the expression levels of key proteins in the cGAS-STING signaling pathway,and ELISA was used to measure the concentrations of interferon-γ(IFN-γ)and tumor necrosis factor-α(TNF-α).The cells in each group were cocultured with MHCC-97H and Huh-7 cells for 6 hours,and then CCK8 assay was used to measure the survival rate of HCC cells in each group.A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups,the Dunnett's T3-test and the Tukey test were used for further comparision between two groups.Results Flow cytometry showed that the purity of γδT cells reached above 99%.Western blot showed that there was no significant difference in the expression of cGAS between the γδT group and the other two groups;compared with the γδT group,the γδT-G10 group had significant increases in the expression levels of STING,phosphorylated STING,TBK1,phosphorylated TBK1,interferon regulatory factor 3(IRF3),and phosphorylated IRF3,while the γδT-H-151 group had significant reductions in the expression of these proteins.ELISA showed that compared with the γδT group,the γδT-G10 group had significant increases in the secretion of IFN-γ and TNF-α by γδT cells(P<0.01 and P<0.05),while the γδT-H-151 group had significant reductions in IFN-γ and TNF-α(P<0.01 and P<0.000 1).CCK-8 assay showed that compared with the γδT group,the γδT-G10 group had significant reductions in the survival rates of the two HCC cell lines(P<0.000 1),while the γδT-H-151 group showed significant increases(P<0.000 1).Conclusion The cGAS-STING signaling pathway can regulate the cytotoxicity of γδT cells against HCC in vitro.关键词
癌,肝细胞/鸟苷酸激酶类/腺苷一磷酸/干扰素调节因子1/T细胞,γ-δKey words
Carcinoma,Hepatocellular/Guanylate Kinases/Adenosine Monophosphate/Interferon Regulatory Factor-1/T-Cell,gamma-delta引用本文复制引用
胡帅,张示杰,王二强,多小勇,徐志,张玉梦,谢士伟,荣利华,王宇晨,李江..环磷酸鸟苷-腺苷合成酶-干扰素基因刺激因子(cGAS-STING)信号通路激活对γδT细胞杀伤肝癌细胞效应的调控作用[J].临床肝胆病杂志,2025,41(12):2572-2580,9.基金项目
国家卫生健康委中亚高发病防治重点实验室开放基金项目(KF202203) （KF202203）
兵团指导性科技计划项目(2023ZD023) （2023ZD023）
院级科技计划项目(BS202203) Open Research Fund of the Key Laboratory of Prevention and Treatment of High Incidence of Central Asia,NHC(National Health Commission)(KF202203) （BS202203）
Guided Science and Technology Plan Project of XPCC(Xinjiang Production and Construction Corps)(2023ZD023) （Xinjiang Production and Construction Corps）
Hospital-Level Science and Technology Plan Project(BS202203) （BS202203）
