Effects of Asiaticoside on Epithelial-Mesenchymal Transition and Immune Escape in Lung Cancer Cells by Regulating the TIM3/Ga19 Axis
This study aims to investigate the effects of ATS(Asiaticoside)on epithelial-mesenchymal transition and immune escape in lung cancer cells by regulating the TIM3(T cell immunoglobulin domain and mucin domain-3)/Ga19(galectin-9)signaling axis.Cells were treated with different concentrations of ATS(0,5,10,20,40,80 μmol/L)to screen for the optimal concentration.The cells were separated into control group,ATS group(25 μmol/L ATS),ATS+pcDNA-NC group(25 μmol/L ATS+pcDNA-NC),and ATS+pcDNA-TIM3 group(25 μmol/L LATS+pcDNA-TIM3).The CCK-8 assay kit and flow cytometry were used to detect cell prolifera-tion and apoptosis.Transwell chamber was used to detect cell invasion and migration.ELISA was applied to de-tect the levels of TNF-α,IFN-γ,and IL-2 in cells.Western blot was applied to detect the expression of E-cadherin,Vimentin,PD-L1,TIM3,and Ga19 proteins in cells.BALB/c nude mice were used to construct NSCLC(non-small cell lung cancer)models,which were separated into control group,ATS group,ATS+AAV-NC group,and ATS+AAV-TIM3 group.Tumor volume and mass were measured.Immunohistochemical staining was applied to observe the expression of PD-L1 and Gal9 proteins.Compared with the control group,the cell survival rate,cell migration number,cell invasion number,Vimentin,PD-L1,TIM3,and Gal9 protein expression in the ATS group were lower(P＜0.05),the apoptosis rate,TNF-α,IFN-γ,IL-2 levels,and E-cadherin protein expression were higher(P＜0.05).Compared with the ATS+pcDNA-NC group,the cell survival rate,cell migration number,cell invasion number,Vimentin,PD-L1,TIM3,and Ga19 protein expression were higher in the ATS+pcDNA-TIM3 group(P＜0.05),the apoptosis rate,TNF-α,IFN-γ,IL-2 levels,and E-cadherin protein expression were lower(P＜0.05).In vivo tumor experiments showed that compared with the control group,the tumor volume and mass of mice in the ATS group were lower(P＜0.05),and PD-L1 and Ga19 protein expression levels were lower.Compared with the ATS+AAV-NC group,the tumor volume and mass of mice in the ATS+AAV-TIM3 group were higher(P＜0.05),and PD-L1 and Ga19 protein expression levels were higher.ATS may inhibit epithelial-mesenchymal transition and immune escape in lung cancer cells by suppressing the TIM3/Ga19 signaling axis.
AsiaticosideT cell immunoglobulin domain and mucin domain-3galectin 9lung cancer cellsimmune escape
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