Basic FGF and IGF-I Promote Differentiation of Human SH-SY5Y Neuroblastoma Cells in Culture

Abstract

Phorbolester-triggered differentiation of SH-SY5Y neuroblastoma cells requires serum and a prolonged activation of protein kinase C (PKC). Under serum-free conditions development of a mature phenotype requires phorbolester in combination with a member of either the insulin-like growth factor (IGF) or the platelet-derived growth factor family. Here we report that basic and acidic fibroblast growth factor (FGF) and epidermal growth factor, but not nerve growth factor, synergistically potentiate phorbolester-induced differentiation. Alone these factors induced a mitogenic response which varied in magnitude, with basic FGF and IGF-I being the two most potent mitogens. However, a combination of basic FGF and IGF-I induced differentiation as judged by morphology and the increase in growth associated protein (GAP-43) and neuropeptide tyrosine mRNA levels. In contrast to the phenotype obtained in the presence of phorbolester, bFGF and IGF-I-treated SH-SY5Y cells retained their capacity to proliferate. Finally, in these cells, the phosphorylation of the endogenous PKC substrate, myristoylated alanine-rich C-kinase substrate (MARCKS), was slightly increased during several days, suggesting an involvement of PKC in the bFGF and IGF-I-induced differentiation.

Molecular dissection of TrkA signal transduction pathways mediating differentiation in human neuroblastoma cells

“…These data are consistent with the previous observation that TrkA-transfected SH-SY5Y cells demonstrated an increased uptake of thymidine after NGF-stimulation, indicating increased mitotic activity (Lavenius et al, 1995). IGF-I-and basic FGF-di erentiated SH-SY5Y cells do also continue to proliferate (Lavenius et al, 1994). However, our data are in contrast to two reports about TrkA-transfected NB cells, in which NGF treatment led to dramatic reduction of cell growth in the MYCN ampli®ed NB cell line HTLA230 (Matsushima and Bogenmann, 1993) and in SH-SY5Y cells (Poluha et al, 1995).…”

Tyrosine Phosphorylation of Paxillin and Focal Adhesion Kinase during Insulin-like Growth Factor-I-stimulated Lamellipodial Advance

“…Tyrosine-phosphorylated proteins were visualized with enhanced chemiluminescence reagents. (19,20,23). These cells grow in serum-free media, allowing analysis of the effects of IGFs in the absence of other growth factors.…”

“…In SH-SY5Y cells, long-term incubation (i.e. 1-3 days) with IGF-I increases neurite length (19,20), although the mechanism of this effect has not been identified. For this reason, we first sought to perform a detailed characterization of the effect of IGF-I in SH-SY5Y cells.…”

“…Human Neuroblastoma Cells-Several laboratories including ours have used SH-SY5Y human neuroblastoma cells to characterize the effects of IGFs on neuronal cells (19,20,23). These cells grow in serum-free media, allowing analysis of the effects of IGFs in the absence of other growth factors.…”

“…We showed recently that FAK and paxillin are tyrosine-phosphorylated during integrin-mediated cell spreading and neurite formation in human neuroblastoma SH-SY5Y cells (18). Because IGF-I also promotes neurite outgrowth in SH-SY5Y cells (19,20), we were interested in determining the function of FAK and paxillin in the response of SH-SY5Y cells to IGF-I. We began our investigations by characterizing the morphological effects of IGF-I on SH-SY5Y cells in detail.…”