半夏泻心汤含药肠吸收液通过干预TA-BMSCs与PMN-MDSCs串话抑制胃癌细胞侵袭迁移OA

Objective:To investigate the synergistic promotion of malignant phenotypes in gastric cancer cells by tumor-associated bone marrow mesenchymal stem cells(TA-BMSCs)and polymorphonuclear myeloid-derived suppressor cells(PMN-MDSCs)through crosstalk,and the intervention mechanism of the intestinal absorption solution containing Banxia Xiexintang(BXT).Methods:Gastric cancer MFC cells were treated with conditioned medium(co-culture-CM)prepared from a co-culture system of TA-BMSCs and PMN-MDSCs.Groups included control,co-culture-CM model,stromal cell-derived factor 1(SDF1)inhibitor(LY2510924),α4β1 inhibitor(BIO5192),dual inhibitor combination,and different concentrations(55%,70%,85%)of BXT-containing intestinal absorption solutions.MFC cell proliferation,migration,invasion,and apoptosis were assessed via cell counting kit-8(CCK-8)assay,wound-healing assay,Transwell assay,and flow cytometry.The SDF-1,α4β1,matrix metalloproteinase-9(MMP-9),and vascular endothelial growth factor A(VEGFA)levels in culture supernatants,along with the protein levels of intracellular macrophage migration inhibitory factor(MIF),chemokine(C-X-C motif)receptor 4(CXCR4),CD106,MMP-9,and VEGFA in MFC cells,were measured by enzyme-linked immunosorbent assay(ELISA)and Western blot,respectively.Results:Compared with the control group,co-culture-CM promoted the proliferation,migration,and invasion of MFC cells(P<0.01),elevated the levels of SDF-1,α4β1,MMP-9,and VEGFA(P<0.01),and upregulated the protein levels of MIF(P<0.05),MMP-9(P<0.01),and VEGFA(P<0.01).Compared with co-culture-CM,BXT-containing intestinal absorption solutions at various concentrations significantly reversed the phenotypic effects on cells,inhibited malignant phenotypes,lowered the levels of MMP-9 and SDF1,and reduced the expression of proteins in the crosstalk axis.Compared with the SDF1+α4β1 inhibitor group,the SDF1 inhibitor group and the α4β1 inhibitor group showed no differences in the inhibition of proliferation,scratch healing,and cytokine levels.The SDF1 inhibitor increased the apoptosis rate and downregulated the protein levels of MIF and CD106,while the α4β1 inhibitor increased the number of migrated cells and the expression of various proteins.Compared with the SDF1 inhibitor group,the α4β1 inhibitor group showed reduced inhibitory effect on proliferation,decreased apoptosis rate,increased number of invasive cells,decreased α4β1 content,and increased expression of various proteins(P<0.01).The 55%,70%,and 85%intestinal absorption solutions increased the inhibitory effect on proliferation,decreased the number of invasive cells,and increased the apoptosis rate(P<0.01).The 55%BXT-containing intestinal absorption solution group showed increased wound healing rate and upregulated protein levels of VEGFA,CD106,and MMP-9(P<0.05).The 70%intestinal absorption solution group showed upregulated protein level of MIF(P<0.05),and the 85%intestinal absorption solution group showed upregulated protein level of VEGFA(P<0.01)and downregulated the protein level of CXCR4(P<0.01).Conclusion:TA-BMSCs and PMN-MDSCs synergistically activate the MIF/SDF-1/CXCR4 and MMP-9/α4β1/CD106 axes through crosstalk,significantly enhancing gastric cancer cell invasion and migration.BXT-containing intestinal absorption fluid effectively inhibits the malignant progression of gastric cancer cells by multi-targeted intervention in this crosstalk process.