Fig. 6

TNF-α and IL-1β promote the expression of OPN in tumor cells and macrophages. A TNF-α and IL-1β activated NF-kappa B signaling pathway in tumor cells. B-E Growth curves and colony formation and migration abilities of HN6 cells stimulated by TNF-α, IL-1β and NF-kappa B inhibitor, PDTC. MTT assays (B), colony formation assays (C), Transwell assays (D) and wound healing assays (E) were performed. Scale bar, 50 μm. F Flow Cytometry showed that TNF-α and IL-1β promoted the expression of OPN in tumor cells. G PCR was used to measure the fold change of OPN in mRNA level in HN6. H PDTC inhibited OPN expression in tumor cells. Relative level was calculated by ImageJ. I TNF-α and IL-1β promoted the expression of OPN in macrophages, which was determined by FCS. J TNF-α and IL-1β activated STAT3 to increase the expression of OPN in macrophages. K, L STAT3 inhibitor, Stattic, inhibited the elevated level of OPN caused by TNF-α and IL-1β in macrophages, both in mRNA (K) and protein (L) level. (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)