Impact of cGAS/STING signaling pathway on biological behaviours of colon cancer cells
Objective To observe the impact of activation or inhibition of cGAS/STING signaling pathway on proliferation,migration and apoptosis of colon cancer cells,and to investigate the role of cGAS/STING signaling pathway in the occurrence of colon cancer.Methods The colon cancer HT29 cells in logarithmic growth phase were divided into the control group(no treatment),STING agonist group(treated with STING agonist-17)and STING inhibitor group(treated with STING inhibitor STING-IN-2).The cells in three groups were cultured in DMEM medium.After 6-h culture,the optical density(OD)value of cell proliferation was detected by CCK-8 assay,the migration rate was detected by cell scratch assay,the number of invasive cells was detected by Transwell chamber assay,the apoptosis rate was detected by flow cytometry,the relative expressions of phosphorylated TANK-binding kinase 1(p-TBK1)and phosphorylated interferon regulatory factor 3(p-IRF3)proteins were detected by Western blot,and the relative expressions of interferon-β(IFN-β),tumor necrosis factor-α(TNF-α),interleukin(IL)-1,IL-6,IL-10 and CXC chemokine ligand 10(CXCL10)mRNAs were detected by real-time fluorescence quantitative PCR.Results The OD value,cell apoptosis rate,migration rate and number of invasive cells showed significant differences in three groups(F=74.527-282.266,all P values＜0.05).The OD values(0.78±0.03,0.46±0.04,0.26±0.04),migration rates[(53.28±2.80)％,(33.27±2.38)％,(16.51±1.67)％],and numbers of invasive cells(322.00±26.63,214.33±22.01,118.33±7.77)decreased sequentially in the STING inhibitor group,control group and STING agonist group(P＜0.05),and the cell apoptosis rates[(5.59±0.59)％,(10.31±0.89)％,(16.79±0.56)％]increased sequentially(P＜0.05).There were significant differences in the relative expressions of p-TBK1 and p-IRF3 proteins,and IFN-β,TNF-α,IL-1,IL-6,IL-10 and CXCL10 mRNAs in three groups(F=321.380-6 276.453,all P values＜0.05).The levels of p-TBK1 protein(0.10±0.01,0.65±0.03,0.99±0.04),p-IRF3 protein(0.21±0.02,0.37±0.02,0.68±0.01),IFN-β mRNA(0.75±0.02,1.00±0.04,2.89±0.04),TNF-α mRNA(0.57±0.03,1.00±0.09,2.85±0.18),IL-1 mRNA(0.59±0.02,1.00±0.02,2.69±0.03),IL-6 mRNA(0.62±0.02,1.00±0.05,2.12±0.05),IL-10 mRNA(0.54±0.01,1.00±0.26,3.71±0.14),and CXCL10 mRNA(0.68±0.02,1.00±0.02,1.93±0.06)increased sequentially in the STING inhibitor group,control group and STING agonist group(P＜0.05).Conclusion To activate the cGAS/STING signaling pathway can inhibit the proliferation,migration and invasion of colon cancer cells,promote the cell apoptosis,and negatively regulate the occurrence and progression of colon cancer,probably by inducing cell apoptosis via inducing the expressions of interferon and inflammatory cytokines.
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