Inhibition of IGF-I–related intracellular signaling pathways by proinflammatory cytokines in growth plate chondrocytes

Abstract

Children with chronic inflammatory diseases suffer from severe growth failure associated with resistance toward the anabolic action of insulin-like growth factor I (IGF-I). We hypothesized that proinflammatory cytokines interfere with IGF-I signaling. We used the mesenchymal chondrogenic cell line RCJ3.1C5.18 as a model of the growth plate. Cell proliferation was assessed by [(3)H]-thymidine-uptake and differentiation by gene expression (quantitative reverse-transcriptase PCR) of specific differentiation markers. Key signaling molecules of the respective IGF-I-related intracellular pathways were determined by western immunoblotting. Coincubation of the proinflammatory cytokines interleukin (IL)-1β (10 ng/ml), IL-6 (100 ng/ml), or tumor necrosis factor-α (50 ng/ml) with IGF-I inhibited IGF-I-driven cell proliferation by 50%, while baseline cell proliferation was not altered. These cytokines attenuated the IGF-I-induced phosphorylation of AKT as a key signaling molecule of the phosphatidylinositol-3 kinase pathway by 30-50% and the phosphorylation of ERK as a key signaling molecule of the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway by 50-75%. Also, IGF-I-enhanced chondrocyte differentiation was inhibited by these proinflammatory cytokines. The insensitivity toward the anabolic action of IGF-I in the growth plate in conditions of chronic inflammation is partially due to inhibition of IGF-I-specific signaling pathways by proinflammatory cytokines, which affect both IGF-I-driven chondrocyte proliferation and differentiation.