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目的 分析车叶草苷酸(ASPA)调节环鸟苷酸-腺苷酸合成酶(cGAS-STING)-干扰素(IFN)基因刺激因子(STING)信号通路对胰腺癌(PC)细胞免疫逃逸的影响。方法 用细胞计数(CCK)-8法分析不同浓度ASPA处理sw1990细胞增殖情况和半抑制浓度(IC50)。将细胞分为标准组、ASPA低、中、高剂量组(L-、M-、H-ASPA组)及RU.521组(cGAS抑制剂RU.521)。观察细胞克隆、增殖及凋亡，并检测自然杀伤(NK)细胞迁移数及杀伤活性。用酶联免疫吸附试验(ELISA)分析共培养液中IFN-γ、粒酶(Granzyme)B表达，Western印迹法检测cGAS、STING、TANK结合激酶(NAK/TBK)1、IFN调节因子(IRF)3、程序性死亡配体(PD)-L1蛋白表达。结果 与标准组比较，L-、M-及H-ASPA组细胞凋亡率、cGAS、STING、NAK/TBK1、IRF3蛋白、NK细胞杀伤活性、IFN-γ、GranzymeB表达明显增加，存活率、克隆细胞数、细胞迁移数、PD-L1蛋白表达明显降低(P<0.05)。与H-ASPA组比较，RU.521组存活率、克隆细胞数、细胞迁移数、PD-L1蛋白表达明显增加，细胞凋亡率、cGAS、STING、NAK/TBK1、IRF3蛋白、NK细胞杀伤活性、IFN-γ、GranzymeB表达明显降低(P<0.05)。结论 ASPA可能参与激活cGAS-STING信号通路，抑制PC细胞免疫逃逸。
Abstract：Objective To analyze the effect of asperulosidic acid(ASPA) on the immune escape of pancreatic cancer(PC) cells by regulating the cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS-STING)-interferon(IFN) gene-stimulating factor(STING) signaling pathway.Methods The cell counting kit(CCK)-8 was used to analyze the proliferation of sw1990 cells treated with different concentrations of ASPA and to determine the half-inhibitory concentration(IC50).The cells were divided into the standard group, the low, medium and high dose groups of ASPA(L-,M-and H-ASPA groups),and the RU.521 group(cGAS inhibitor RU.521).The cell clone, proliferation and apoptosis were observed, and the migration number and killing activity of natural killer(NK) cells were detected.The enzyme-linked immunosorbent assay(ELISA) was used to analyze the expressions of IFN-γ and GranzymeB in the co-culture medium.Western blotting was used to detect the expressions of cGAS,STING,TANK-kinase(NAK/TBK) 1,IFN regulatory factor(IRF)3 and programmed death ligand(PD)-L1 protein.Results Compared with the standard group, the apoptosis rate, cGAS,STING,NAK/TBK1,IRF3 protein, NK cell killing activity, IFN-γ and GranzymeB expressions were significantly increased, while the survival rate, the number of cloned cells, the number of cell migrations and PD-L1 protein expression were significantly decreased in the L-,M-,and H-ASPA groups(P<0.05).Compared with the H-ASPA group, the survival rate, the number of cloned cells, the number of cell migrations and the expression of PD-L1 protein were significantly increased, while the apoptosis rate, cGAS,STING,NAK/TBK1,IRF3 protein, NK cell killing activity, IFN-γ and GranzymeB expressions were significantly decreased in the RU.521 group(P<0.05).Conclusions ASPA may participate in activating the cGAS-STING signaling pathway and thereby inhibit immune escape by PC cells.
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基本信息:
中图分类号:R735.9
引用信息:
[1]丁世林,汤杰,陈建林,等.车叶草苷酸激活cGAS-STING信号通路抑制胰腺癌细胞免疫逃逸[J].中国老年学杂志,2026,46(06):1060-1065.
基金信息:
国家自然科学基金面上项目(82172931,32170915); 如皋市科技攻关计划项目(SRGS(23)020)