Research Paper Volume 13, Issue 17 pp 21571—21586

Figure 5. TNFSF9 induces M2 polarization of macrophages and promotes the migration of pancreatic cancer cells. (A) U937 cells were successfully induced into macrophages. (B, C) The effect of wild-type pancreatic cancer cells on the polarization of U937-derived macrophages. (D, E) TNFSF9 knockdown pancreatic cancer cells inhibit the expression of M2 markers (IL-10 and TGF-β) mRNA levels in U937 macrophages. At the same time, it promotes the expression of M1 marker (IL-8 and TNF-α) mRNA level and inhibits the expression of IL-1β mRNA level. (F, G) Co-culture of U937-derived macrophages with TNFSF9 knockdown pancreatic cancer cells inhibited the migration of BXPC-3 and PANC-1 cells. (H, I) The migration of BXPC-3 and PANC-1 cells increased after adding recombinant human protein TGF-β. (J–L) Western blots were used to analyze the expression of inflammatory cytokines IL-8, IL-6, TNF-α and IL-1β on U937-derived macrophages after co-culture with pancreatic cancer cells knocked down by TNFSF9. Compared with the sh-NC group, the expression of IL-8, IL-6 and TNF-α in the sh-TNFSF9-1 and sh-TNFSF9-2 groups were significantly increased, and the expression of IL-1β was significantly reduced. sh-1 is sh-TNFSF9-1. sh-2 is sh-TNFSF9-2. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 and ^#P < 0.001.