title: Lysophosphatidic acid-induced ERK activation and chemotaxis in MC3T3-E1 preosteoblasts are independent of EGF receptor transactivation.
abstract: Bone-forming osteoblasts and their progenitors are target cells for the lipid growth factor lysophosphatidic acid (LPA) which is produced by degranulating platelets at sites of tissue injury. LPA is a potent inducer of bone cell chemotaxis, proliferation and survival in vitro, and this lipid factor is an attractive candidate to facilitate preosteoblast migration during skeletal regeneration in vivo. In this study we sought to more clearly define the intracellular signaling pathways mediating the effects of LPA on bone cells. LPA-treated MC3T3-E1 preosteoblastic cells exhibited a bimodal activation of extracellular signal-related kinase (ERK1/2) with maximal phosphorylation at 5 and 60 min. MEK1/2 activation was detected within 2.5 min of LPA exposure and remained elevated for at least an hour. ERK1/2 phosphorylation was not coupled to Ras activation or to LPA-induced elevations in cytosolic Ca(2+). While LPA exposure transactivates the EGF receptor in many cell types, LPA-stimulated ERK1/2 activation in MC3T3-E1 cells was unaffected by the inhibition of EGF receptor function. ERK isoforms can function as transcription factors and ERK1/2 rapidly accumulated in the nuclei of LPA-treated cells, a process that was blocked if ERK1/2 phosphorylation was prevented. Blocking ERK1/2 phosphorylation also led to significant decreases in LPA-induced MC3T3-E1 cell chemotaxis, while the inhibition of EGF receptor function had no effect on the stimulation of preosteoblast motility by LPA. Our results identify ERK1/2 activation as a mediator of LPA-stimulated MC3T3-E1 cell migration that may be relevant to preosteoblast motility and gene expression during bone repair in vivo. J. Cell. Physiol. 219: 716-723, 2009. (c) 2009 Wiley-Liss, Inc.
authors.affiliation: Cell Biology and Biochemistry Group, Pacific Northwest National Laboratory, Richland, Washington.
authors.forename: Sue A
authors.initials: SA
authors.lastname: Karagiosis
authors.forename: William B
authors.initials: WB
authors.lastname: Chrisler
authors.forename: Nikki
authors.initials: N
authors.lastname: Bollinger
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Karin
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002465:Cell Movement / Q000187:drug effects; D002633:Chemotaxis / Q000187:drug effects* / Q000502:physiology; D017931:DNA Primers / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects; D066246:ErbB Receptors / Q000235:genetics*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D008246:Lysophospholipids / Q000494:pharmacology*; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D010006:Osteoblasts / Q000187:drug effects* / Q000502:physiology*; D010766:Phosphorylation; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection
chemical_list: D017931:DNA Primers; D008246:Lysophospholipids; D066246:ErbB Receptors; D048049:Extracellular Signal-Regulated MAP Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; C032881:lysophosphatidic acid