Figure 4.

Changes in glucose metabolism, cytokine and chemokine expression, and JAK/STAT3 signaling seen with co-culture of cervical cancer cells and macrophages and macrophage-conditioned media. Co-culture of macrophages with ME180 and SiHa cells, or culturing ME180 and SiHa cells with macrophage-conditioned media, results in a significant increase in 2-NBDG signal measured by flow cytometry; this is most pronounced in the SiHa cells (A). Differentiated macrophages produce a number of cytokines; co-culture with ME180 cells significantly increased expression of IL-1β and IL-6 compared to differentiated macrophages alone (B). Co-culture with differentiated macrophages, or culturing with macrophage-conditioned media, induces STAT3 phosphorylation in ME180 cells, which have little STAT3 phosphorylation at baseline. This effect was not seen in SiHa cells, which show high baseline levels of STAT3 phosphorylation (C). SiHa cells cultured with IL-6 NAB do show reduced STAT3 phosphorylation (D). Treatment of macrophages co-cultured with cervical cancer cells with IL-6 NAB alters 2-NBDG uptake in both ME180, and SiHa cells. This effect was most pronounced with SiHa cells, and co-culture of either SiHa or ME180 cells with M2 macrophages (E). GSEA analysis showed that the Hallmark Glycolysis pathway was significantly upregulated in tumors with high STAT3 expression (F). Experiments were carried out in triplicate (n = 3) with error bars representing standard deviation.