Effect of Germacrone on Epithelial Mesenchymal Transition and Immune Escape in Gastric Cancer Cells by Regulating the STING/TBK1/IRF3 Signaling Pathway
ZHAI Haoqing1*, CHENG Lin1, CHEN Lan2
This study aims to investigate the effect of GM (germacrone) on epithelial mesenchymal transition and immune escape of GC (gastric cancer) cells by regulating the STING (stimulator of interferon gene)/TBK1 (TANK binding kinase 1)/IRF3 (interferon regulatory factor 3) signaling pathway. Human GC cells (BGC-823) cultured in vitro were randomly grouped into Control group (normal culture), L-GM, M-GM, H-GM groups (20, 40, 80 μmol/L GM), and C-176 group (80 μmol/L GM+1 μmol/L STING/TBK1/IRF3 signaling pathway inhibitor C-176). Cell proliferation was detected by CCK8 and colony formation assay. Cell migration was determined by a scratch assay, while cell invasion and apoptosiswere evaluated by Transwell and flow cytometry assays, respectively. Western blot was used to detect the expression of Vimentin, E-cadherin, PD-L1, STING/TBK1/IRF3 signaling pathway related proteins (STING, p-TBK1/TBK1, p-IRF3/ IRF3) in cells. After coculture BGC-823 cells and CD8+ T cells, CD8+ T cells were separated into coculture group, GM group, and inhibitor group, and the killing rate of activated CD8+ T cells was calculated. Compared with the Control group, the survival rate, clone number, scratch healing rate, invasion number, PD-L1, and Vimentin protein expression of BGC-823 cells in the L-GM group, M-GM group, and H-GM group were lower, while the apoptosis rate, E-cadherin, STING, p-TBK1/TBK1, and p-IRF3/IRF3 expression were higher (P<0.05). Compared with the H-GM group, the survival rate, clone number, scratch healing rate, invasion number, PD-L1, and Vimentin protein expression of cells in the C-176 group were higher, while the apoptosis rate, E-cadherin, STING, p-TBK1/TBK1, and p-IRF3/IRF3 expression were lower (P<0.05). Compared with the coculture group, the killing rate of activated CD8+ T cells in the GM group higher (P<0.05). Compared with the GM group, the killing rate of activated CD8+ T cells was lower in the inhibitor group (P<0.05). GM inhibits epithelial mesenchymal transition, cell proliferation, migration, invasion, and GC immune escape in BGC-823 cells by regulating the STING/TBK1/IRF3 signaling pathway.
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