Fig. 9

Bone marrow stromal cells are the primary contributors of production of IL-6 and IL-8 in co-cultivation with MCF-7 breast cancer cells. ELISA assays demonstrated significantly higher levels of production of murine (a). IL-6 and (b). IL-8. MCF-7 breast cancer cells (1000 cells/well) were seeded on human fibronectin coated 24-well plate. On day 3, cells were treated with H₂O₂ and CCCP for one hour in serum-free medium, and with ICI for 2 days in DMEM/10%FCS medium without phenol-red, washed once and incubated in DMEM/10%FCS. Confluent stromal monolayers were prepared from murine bone marrow stroma as before. At confluence, stromal monolayers were treated with H₂O₂ and CCCP for one hour and with ICI for 2 days, washed once and incubated in DMEM/10%FCS medium. MCF-7 breast cancer cells (1000 cells/well) were co-incubated with control and H₂O₂ 50 μM, CCCP 50 μM and ICI-treated stromal monolayers for 6 days. Supernatant samples were collected 24, 48 and 120 h and murine and human IL-6 and IL-8 levels in the supernatant were determined using murine and human IL-6 ELISA kit, and murine CXCL1/KC and human CXCL8/IL-8 ELISA kit according to the manufacturer’s instructions. Error bars: Standard Deviation. *p < 0.05, **p < 0.01 ***p < 0.001