Effect Of IL-8 Derived From Gastric Mesenchymal Stem Cells On Migration Of Gastric Cancer Cells And Its Mechanism

Objective: This study aimed to analyze the role of IL-8 derived from gastric cancer mesenchymal stem cells on migration and invasion of gastric cancer cell lines,and the potential molecular mechanism,finding potential molecular targets for the prevention and treatment of treatment of gastric cancer and to provide theoretical basis.Methods: GC-MSCs and BM-MSCs were isolated and cultured from the tumor tissues of gastric cancer patients and normal human bone marrow by tissue cell adherence method,and gastric cancer cell lines(SGC-7901)were cultured.Their culture medium(GC-MSCs-CM,BM-MSCs-CM and SGC-7901-CM)were collected.Enzyme-linked immunosorbent assay(Elisa)to detect the expression level of IL-8 in GC-MSCs-CM,BM-MSCs-CM and the culture medium of gastric cancer cell line(SGC-7901).In order to verify the effect of GC-MSC on GC migration,BM-MSCs-CM and GC-MSCs-CM were applied to gastric cancer cell lines(SGC-7901).We use Transwell assay and cell wound scratch assay to detect the migration ability of gastric cancer cells.In order to verify the role of IL-8 in GC-MSCs-CM to promote the progress of GC,the IL-8 antibody was used to neutralize IL-8 in GC-MSCs-CM and then applied to gastric cancer cell line(SGC-7901).Cell wound scratch assay to detect the migration ability of gastric cancer cells,Western-blotting detection of gastric cancer cell-associated epithelial-mesenchymal transition marker proteins E-cadherin,N-cadherin,Vimentin expression level.In order to explore the signaling pathway of IL-8 from GC-MSCs to SGC-7901 to release matrix metalloproteinase 9(MMP-9),Western-blotting was used to detect the expression levels of MMP-9,Akt,and Erk signaling pathway-related proteins and phosphorylation,and through exogenous IL-8 treatment of gastric cancer cells(SGC-7901)validated the signaling pathway.In order to evaluate the effect of IL-8 on the survival rate of gastric cancer patients,We analyzed the relationship between IL-8 mRNA expression and the overall survival rate of gastric cancer patients was analyzed through the Kmplot databaseResults: Elisa test results showed that IL-8 expression was higher than that of bone marrow mesenchymal stem cell supernatant and gastric cancer tissue supernatant than gastric cancer mesenchymal stem cell supernatant(P <0.001).Gastric cancer mesenchymal stem cells can promote the migration of gastric cancer cell line SGC-7901;after neutralizing IL-8 in GC-MSCs-CM,the migration ability of gastric cancer cell line SGC-7901 is weakened,and the expression changes of EMT-related proteins are vimentin and The expression of N-cadherin decreased,the expression of E-cadherin increased,inhibited the occurrence of EMT process,and after neutralizing IL-8 GC-MSCs-CM treatment of SGC-7901,Transwell experiment found that the number of cells migrated through the membrane was obvious Decreased,the cell scratch test found that the cell healing ability was significantly weakened,inhibiting the migration ability of gastric cancer cells;after neutralizing IL-8 in GC-MSCs-CM,PI3 K / Akt and MAPK / Erk signaling pathway activation was weakened,and MMP-9 protein expression decreased,inhibiting the release of MMP,suggesting that GC-MSCs may promote the expression of MMP-9 through PI3 K / Akt and MAPK / Erk signaling pathways,thereby promoting gastric cancer invasion.Recombinant human IL-8 protein promotes the expression of PI3 K / Akt,MAPK / Erk signaling pathway and MMP-9.Patients with high expression of IL-8mRNA have low overall survival rate,especially in patients with lymph node metastasis or pathological type of diffuse gastric cancer.Conclusion: IL-8,derived from GC-MSCs-CM,plays an key role in promoting gastric cancer migration and invasion.It may be that GC-MSCs-CM regulates gastric cancer EMT and promotes high expression of MMP-9 through PI3 K / AKT and MAPK / Erk signaling pathways to promote gastric cancer progression.IL-8 may be one of the significant factors regulating the migration and invasion of gastric cancer cells,which provides a preliminary experimental basis for further exploring the mechanism of GC-MSCs promoting tumor invasion and exploring potential targets.