苍术素调节cGAS-STING信号通路介导的免疫反应对肺癌细胞恶性生物学行为的影响

The Effect of Atractylodin on the Malignant Biological Behavior of Lung Cancer Cells by Regulating the Immune Response Mediated by cGAS-STING Signal Pathway

目的探讨苍术素(ATR)调节环磷酸鸟苷-腺苷酸合成酶(cGAS)-干扰素基因刺激蛋白(STING)信号通路介导的免疫反应对肺癌细胞恶性生物学行为的影响。方法以不同浓度的ATR及不同的作用时间处理A549细胞,采用CCK-8法筛选最佳的ATR浓度及作用时间。取生长状态较好的A549细胞,分为空白组、对照组(0.1%二甲基亚砜)、ATR组(40mg/L ATR)、ATR+siRNA敲降cGAS(si cGAS)组(40mg/L ATR+转染si cGAS)、ATR+阴性对照(si NC)组(40mg/L ATR+转染si NC)。48h后,流式细胞仪、CCK-8分别检测细胞凋亡及增殖变化;Transwell实验检测细胞迁移、侵袭;Western blot检测细胞中金属基质蛋白酶-9(MMP-9)、凋亡蛋白(Bax)、增殖蛋白(CyclinD1)、cGAS、STING蛋白表达。结果40mg/L ATR作用48h为最佳的处理A549细胞的浓度及时间。与空白组、对照组相比,ATR组细胞侵袭及迁移数、增殖率、CyclinD1、MMP-9表达显著下降,凋亡率、cGAS、STING、Bax表达显著增加(P<0.05);与ATR+si NC组相比,ATR+si cGAS组细胞侵袭及迁移数、增殖率、CyclinD1、MMP-9表达显著增加,凋亡率、cGAS、STING、Bax表达显著降低(P<0.05)。结论ATR可通过激活cGAS-STING信号通路介导的免疫反应抑制肺癌细胞恶性生物学行为。

Objective To investigate the effect of atractylodin(ATR)on the malignant biological behavior of lung cancer cells by the cyclic guanosine monophosphate adenylate synthetase(cGAS)-stimulator of interferon gene(STING)signal pathway mediated immune response.Methods A549 cells were treated with different concentrations of ATR and different action time points.CCK-8 method was used to select the optimal concentration and action time points of ATR.A549 cells in the good growth condition were divided into the blank group,control group(0.1%dimethyl sulfoxide),ATR group(40 mg/L ATR),ATR+siRNA knockdown cGAS(si cGAS)group(40 mg/L ATR+transfected with si cGAS),and ATR+negative control(si NC)group(40 mg/L ATR+transfected with si NC).After 48h,cell apoptosis and proliferation were detected respectively by flow cytometry and CCK-8;Transwell assay was used to detect cell migration and invasion;Western blot was used to detect the expression of matrix metalloproteinase-9(MMP-9),apoptosis protein(Bax),proliferating protein(CyclinD1),and cGAS,STING protein in cells.Results 40mg/L ATR for 48h was the optimal concentration and time point of A549 cells.Compared with the blank group and control group,the numbers of cell invasion and migration,proliferation rate,expression of cyclinD1 and MMP-9 in ATR group decreased significantly,while the apoptosis rate,expression of cGAS,STING and Bax increased significantly(P<0.05);Compared with ATR+si NC group,the numbers of cell invasion and migration,proliferation rate,expression of cyclinD1 and MMP-9 in ATR+si cGAS group increased significantly,while the apoptosis rate,expression of cGAS,STING and Bax decreased significantly(P<0.05).Conclusion ATR can inhibit the malignant biological behavior of lung cancer cells by activating the cGAS-STING signal pathway mediated immune response.