A clinicopathological study on correlation between CD74+fibroblasts and the formation of immunosuppressive microenvironment in oral squamous cell carcinoma
Objective:To investigate the expression of CD74+fibroblasts in oral squamous cell carcinoma(OSCC)and their correlation with the immunosuppressive microenvironment of OSCC.Methods:The expression of CD74 of fibroblasts in OSCC tumor tissues and adjacent non-cancerous tissues will be detected using immunohistochemistry.The correlation between the expression of CD74 in cancer-associated fibroblasts(CAFs)and patients'clinicopathological parameters,prognosis,and immune cell infiltration will be further investigated.Primary CAFs,paired normal fibroblasts(NFs),and human peripheral blood mononuclear cells(PBMCs)from non-OSCC patients will be extracted to compare the differences in the expression of CD74 between the two types of fibroblasts.Subsequently,these two types of fibroblasts will be co-cultured with PBMCs,and the concentrations of interleukin-10(IL-10)and IL-35 in the culture supernatant will be measured by enzyme-linked immunosorbent assay(ELISA).The effects of CAFs with different CD74 expression levels on the migration of PBMCs were compared by Transwell assay and flow cytometry.Results:The number of CD74+fibroblasts in OSCC tumor tissues was significantly higher than that in adjacent non-cancerous tissues(P<0.001).Moreover,OSCC patients with high CD74 expression had a significantly lower 5-year overall survival rate(P<0.001).Immunohistochemical results showed that in tumor tissues with high CD74+fibroblast expression,the ratio of FOXP3+cells to CD4+cells was significantly higher(P<0.001),and there was a significant positive correlation between the number of CD74+fibroblasts and FOXP3+cells(r=0.439 3).In vitro experiments confirmed that the expression of CD74 in primary CAFs from OSCC tumor tissues was higher than that in the paired NFs.After co-culturing with PBMCs for 12 h,the concentrations of IL-10 and IL-35 in the culture medium of the CAFs CD74 high-expression group were significantly higher than those in the NFs group(P<0.05).Additionally,the concentrations of IL-35 in the culture medium of the CAFs CD74 high-expression group were higher than those in the CAFs CD74 low-expression group(P<0.05).The results of flow cytometry showed that CAFs with high CD74 expression had a stronger chemotactic ability toward CD4+FOXP3+cells(P<0.05).Conclusion:CD74+fibroblasts are associated with poorer prognosis in OSCC patients,and may be involved in the recruitment of CD4+FOXP3+cells.
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