Figure 6: The TNF inflammatory pathway affects phagocytosis of breast cancer cells through the regulation of CD47 expression. | Nature Communications

Figure 6: The TNF inflammatory pathway affects phagocytosis of breast cancer cells through the regulation of CD47 expression.

From: A CD47-associated super-enhancer links pro-inflammatory signalling to CD47 upregulation in breast cancer

Figure 6

(a) Flow cytometry analysis showing TNFR1 expression in the breast cancer line MCF7 (CD47Hi), non-tumorigenic breast line MCF10 (CD47Med) and HepG2 (CD47Lo) hepatoma cancer line shows that TNFR1 expression is higher in the MCF7 breast cancer cells. Mean values of TNFR1-FITC fluorescence are shown in blue to the right of each grey histogram. Grey histograms represent FMO controls. (b) Upper figure: A greater increase in CD47 expression is observed in MCF7 when compared to MCF10 or HepG2 cell lines after stimulation with TNF-α (green histogram). Mean values of CD47-APC are shown next to each histogram. Lower figure: Increase in CD47 transcript expression begins to occur after 8 h in MCF7 cells and after 24 h in HepG2 cells upon TNF-α stimulation. N=3 samples. Values represent mean+s.d. Student’s unpaired t-test for independent samples was performed. ***P<0.001. (c) Treating patient-derived xenografted (PDX) and primary patient (PP) breast tumour cells with TNF-α significantly increases CD47 gene expression in four out of five breast tumour samples. Values represent mean+s.d. Student’s unpaired t-test for independent samples was performed. ***P<0.001, **P<0.01, *P<0.1. NS=not significant. ER+ PR+=oestrogen and progesterone positive; TN=triple negative. (d) Upper panels: Blocking TNF-α with infliximab reduces CD47 expression regardless of TNF-α stimulation. Lower panels: Blocking TNF-α with infliximab while knocking down NFKB1 causes a greater reduction on CD47 expression regardless of TNF-α stimulation. (e) Treating MCF7 with infliximab increases phagocytosis of the cancer cells, and, to a greater extent, if infliximab is combined with CD47 blocking antibody. A more pronounced increase in phagocytosis of the cancer cells was observed for MCF7 cells that have NFKB1 knockdown and were treated with the infliximab and CD47 antibodies combined. N=6 samples. Values represent mean+s.d. ****P<0.0005, ***P<0.001, **P<0.05, *P=0.15 (upper figure), *P=0.20 (bottom figure).

Back to article page