麦冬皂苷B调节cGAS-STING信号通路对甲状腺癌TPC-1细胞凋亡、增殖和免疫逃逸的影响OA
Effects of OP-B on apoptosis,proliferation and immune escape of thyroid cancer TPC-1 cells by regulating cGAS-STING signaling pathway
目的:探讨麦冬皂苷B(OP-B)对甲状腺癌TPC-1细胞凋亡、增殖和免疫逃逸的影响及其与环状GMP-AMP合成酶(cGAS)-干扰素基因刺激蛋白(STING)信号通路的调节机制.方法:将TPC-1细胞分为TPC-1组(未做任何处理的 TPC-1 细胞)、L-OP-B 组(2.5 μmol/L OP-B 处理 TPC-1 细胞)、M-OP-B 组(5 μmol/L OP-B 处理TPC-1 细胞)、H-OP-B 组(10 μmol/L OP-B 处理 TPC-1 细胞)、RU.521 组(100μmol/L cGAS-STING 信号通路抑制剂 RU.521 处理 TPC-1 细胞)、H-OP-B+RU.521 组(100 μmol/L RU.521 和 10 μmol/L OP-B 共同处理 TPC-1细胞).MTT法检测OP-B对正常人甲状腺细胞系NTHY ORI 3-1细胞和TPC-1细胞的毒性;MTT法检测TPC-1细胞增殖;流式细胞术检测TPC-1细胞凋亡;将CD8+T细胞与各组处理的TPC-1细胞共培养,台盼蓝染色及流式细胞术检测CD8+T细胞活力;酶联免疫吸附(ELISA)检测上清液中γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)水平;Western blot检测程序性死亡配体1(PD-L1)、cGAS-STING通路蛋白水平.结果:与TPC-1组比较,L-OP-B组、M-OP-B组、H-OP-B组OD值和PD-L1蛋白水平显著降低,CD8+T细胞比例、CD8+T细胞存活率、TPC-1凋亡率、IFN-7和TNF-α水平、cGAS和STING蛋白水平显著升高,且OP-B的作用效果呈现剂量相关性,而RU.521组以上指标呈现相反的趋势(均P＜0.05).与H-OP-B组比较,H-OP-B+RU.521组OD值、PD-L1蛋白水平显著升高,TPC-1凋亡率、CD8+T细胞比例、CD8+T细胞存活率、IFN-γ和TNF-α水平、cGAS和STING蛋白水平显著下降(均P＜0.05).结论:OP-B可诱导TPC-1细胞凋亡,抑制细胞增殖和免疫逃逸,其机制可能与激活cGAS-STING 信号通路相关.
Objective:To investigate the effect of Ophiopogonin B(OP-B)on the apoptosis,proliferation and im-mune escape of thyroid cancer TPC-1 cells and the regulatory mechanism of cyclic GMP-AMP synthase(cGAS)-stimulator of interferon gene(STING)signaling pathway.Methods:TPC-1 cells were grouped into TPC-1 group(un-treated TPC-1 cells),L-OP-B group(2.5 μmol/L OP-B treated TPC-1 cells),M-OP-B group(5 μmol/L OP-B trea-ted TPC-1 cells),H-OP-B group(10 μmol/L OP-B treated TPC-1 cells),RU.521 group(100 μmol/L cGAS-STING signaling pathway inhibitor RU.521 treated TPC-1 cells),and H-OP-B+RU.521 group(100 μmol/L RU.521 and 10 μmol/L OP-B co-treated TPC-1 cells).MTT method was applied to detect the toxicity of OP-B on normal human thyroid cell line NTHY ORI 3-1 cells and TPC-1 cells.MTT method was applied to detect TPC-1 cell proliferation.Flow cytometry was applied to detect apoptosis of TPC-1 cells.CD8+T cells were co-cultured with TPC-1 cells in each group,detection of CD8+T cell viability was performed by trypan blue staining and flow cytometry.ELISA method was applied to detect the levels of IFN-γ and TNF-α in the supernatant.Western blot was applied to detect programmed death ligand 1(PD-L1)and cGAS-STING pathway protein levels.Results:Compared with the TPC-1 group,the OD value and PD-L1 protein level in the L-OP-B group,M-OP-B group,and H-OP-B group were obviously reduced,the apoptosis rate of TPC-1 cells,proportion of CD8+T cells,survival rate of CD8+T cells,the IFN-γ and TNF-α levels,cGAS and STING protein levels were obviously increased,and the effect of OP-B showed a dose-de-pendent relationship,while the RU.521 group showed an opposite trend in the above indicators(all P＜0.05).Com-pared with the H-OP-B group,the OD value and protein level of PD-L1 in the H-OP-B+RU.521 group were obvi-ously increased,the apoptosis rate of TPC-1 cells,proportion of CD8+T cells,survival rate of CD8+T cells,the IFN-γand TNF-α levels,cGAS and STING protein levels were obviously reduced(all P＜0.05).Conclusion:OP-B can in-duce apoptosis of TPC-1 cells,inhibit cell proliferation and immune escape,and the mechanism may be related to the activation of cGAS-STING signaling pathway.
陈攀;张煜;赵启生
湖北医药学院附属随州医院随州市中心医院甲乳外科,湖北随州 441300湖北医药学院附属随州医院随州市中心医院甲乳外科,湖北随州 441300湖北医药学院附属随州医院随州市中心医院甲乳外科,湖北随州 441300
医药卫生
甲状腺癌麦冬皂苷B增殖凋亡免疫逃逸环状GMP-AMP合成酶-干扰素基因刺激蛋白信号通路
Thyroid cancerOphiopogonin BProliferationApoptosisImmune escapeCyclic GMP-AMP syn-thase-stimulator of interferon genes signaling pathway
《陕西医学杂志》 2026 (6)
769-774,6
湖北省卫生健康科研指导性项目(WJ2021F087)
评论