Overexpression of TIA1 gene in M2-type tumor-associated macrophages inhibited invasion and migration of gastric cancer cells by regulating the PI3K/AKT signaling pathway
Objective:To investigate the effect of overexpression of T cell restricted intracellular antigen 1(TIA1)gene in M2-type tumor-associated macrophages(M2-TAMs)on invasion and migration of gastric cancer cells and its mechanism.Methods:Primary TAMs were extracted from gastric cancer tissues and induced to differentiate into M2-TAMs using IL-4 and IL-13.The TIA1 overex-pression plasmid(oe-TIA1)and its empty vector were transfected into M2-TAMs.The expression levels of TIA1 mRNA and protein were detected by qRT-PCR and Western blot.The expression levels of CD206 and CD163 were detected by flow cytometry.The levels of IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant were detected by ELISA.The transfected M2-TAMs were co-cultured with gastric cancer BGC-823 cells by Transwell co-cultivation system,and PI3K agonist 740Y-P was used to intervene in parallel.The cell migration ability was detected by scratch assay.Transwell assay was used to detect cell invasion ability.Protein expression levels of PI3K,p-PI3K,AKT,p-AKT,MMP-2 and MMP-9 in the cells were detected by Western blot.Results:①Compared with primary TAMs,the levels of CD206 and CD163 expression in M2-TAMs cells and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture superna-tant were significantly increased(P<0.05),while the expression levels of TIA1 mRNA and protein were significantly decreased(P<0.05).Overexpression of TIA1 gene significantly decreased the expression levels of CD206 and CD163 in M2-TAMs and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant(P<0.05).②Overexpression of TIA1 gene in M2-TAMs could significantly reduce the migration and invasion ability and the expression levels of p-PI3K/PI3K,p-AKT/AKT,MMP-2 and MMP-9 proteins in BGC-823 cells(P<0.05).③740Y-P could significantly reverse the inhibitory effects of overexpression of TIA1 gene in M2-TAMs on migration,inva-sion and PI3K/AKT signaling pathway of BGC-823 cells.Conclusion:Overexpression of TIA1 gene in M2-TAMs can affect the inva-sion and migration of gastric cancer cells by blocking the PI3K/AKT signaling pathway.
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