title: Fibroblast growth factor-2 over-rides insulin-like growth factor-I induced proliferation and cell survival in human neuroblastoma cells.
abstract: The insulin-like growth factor (IGF) system is a key regulator of cell growth, survival and differentiation, and these functions are co-modulated by other growth factors including fibroblast growth factor-2 (FGF-2). To investigate IGF/FGF interactions in neuronal cells, we employed neuroblastoma cells (SK-N-MC). In serum free conditions proliferation of the SK-N-MC cells was promoted by IGF-I (25 ng/ml), but blunted by FGF-2 (50 ng/ml). IGF-I-induced proliferation was abolished in the presence of FGF-2 even when IGF-I was used at 100 ng/ml. In addition to our previously described FGF-2 induced proteolytic cleavage of IGFBP-2, we found that FGF-2 increased IGFBP-6 levels in conditioned medium (CM) without affecting IGFBP-6 mRNA abundance. Modulation of IGFBP-2 and -6 levels were not significant mechanisms involved in the blockade of IGF-I action since the potent IGF-I analogues [QAYL]IGF-I and des(1-3)IGF-I (minimal IGFBP affinity) were unable to overcome FGF-2 inhibition of cell proliferation. FGF-2 treated cells showed morphological differentiation expressing the TUJ1 neuronal marker while cells treated with IGF-I alone showed no morphological change. When IGF-I was combined with FGF-2, however, cell morphology was indistinguishable from that seen with FGF-2 alone. FGF-2 inhibited proliferation and enhanced differentiation was also associated with a 70% increase in cell death. Although IGF-I alone was potently anti-apoptotic (60% decreased), IGF-I was unable to prevent apoptosis when administrated in combination with FGF-2. Gene-array analysis confirmed FGF-2 activation of the intrinsic and extrinsic apoptotic pathways and blockade of IGF anti-apoptotic signaling. FGF-2, directly and indirectly, overcomes the proliferative and anti-apoptotic activity of IGF-I by complex mechanisms, including enhancement of differentiation and apoptotic pathways, and inhibition of IGF-I induced anti-apoptotic signalling. Modulation of IGF binding protein abundance by FGF-2 does not play a significant role in inhibition of IGF-I induced mitogenesis.
authors.affiliation: Centre for Hormone Research, Parkville, Vic., Australia. russov@cryptic.rch.unimelb.edu.au
authors.forename: Vincenzo C
authors.initials: VC
authors.lastname: Russo
authors.forename: E
authors.initials: E
authors.lastname: Andaloro
authors.forename: S A
authors.initials: SA
authors.lastname: Fornaro
authors.forename: S
authors.initials: S
authors.lastname: Najdovska
authors.forename: D F
authors.initials: DF
authors.lastname: Newgreen
authors.forename: L A
authors.initials: LA
authors.lastname: Bach
authors.forename: G A
authors.initials: GA
authors.lastname: Werther
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D015152:Blotting, Northern; D015153:Blotting, Western; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000187:drug effects; D018976:Insulin-Like Growth Factor Binding Protein 6 / Q000187:drug effects; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis
chemical_list: D018971:Insulin-Like Growth Factor Binding Protein 2; D018976:Insulin-Like Growth Factor Binding Protein 6; D016222:Fibroblast Growth Factor 2; D007334:Insulin-Like Growth Factor I