Synergistic effects of tumor necrosis factor-α and insulin-like growth factor-I on survival of human trophoblast-derived BeWo cell line

Abstract Objective Trophoblast survival is regulated by cytokines and growth factors. While the pharmacological levels (10–100 ng/mL) of tumor necrosis factor (TNF)- α affect trophoblasts survival in vitro, the effects of the physiological levels (1–10 pg/mL) of TNF-α remain unknown. We investigated the effects of the physiological levels of TNF-α on proliferation and apoptosis of human trophoblast cells by using BeWo cells. Insulin-like growth factor (IGF)-I is also a potent regulator of trophoblast survival and has been known to exert synergistic effects with other hormones. The interaction of IGF-I and TNF-α on BeWo cells survival was also examined. Methods After incubating BeWo under the presence of TNF-α (10–10 5  pg/mL) and IGF-I (10 2  ng/mL), we assessed cell number by WST-1 assay and cell proliferation by BrdU uptake assay and immunocytochemistry with anti-Ki67 antibody. Apoptosis was evaluated by TUNEL assay and caspase-3, 8 activity assays. Results Under the presence of IGF–I, cell number, BrdU uptake, and Ki-67 expression of BeWo were dose-dependently enhanced by low TNF-α (10–10 2  pg/mL), while no such effects were detected without IGF–I. Higher levels of TNF-α (10 4 –10 5  pg/mL) showed inhibiting effects on cell number and cell proliferation. The number of TUNEL positive cells were decreased and caspase activities were suppressed by lower levels (10–10 2  pg/mL) of TNF-α and IGF-I independently. Higher levels of TNF-α (10 4 –10 5  pg/mL) showed promoting effects on apoptosis irrespective of IGF–I. Conclusion The physiological levels of TNF-α and IGF-I had synergetic effects on enhancing cell proliferation and also independently inhibited apoptosis of Bewo cells.