Fibroblast Growth Factor-2 Suppression of Tumor Necrosis Factor α-Mediated Apoptosis Requires Ras and the Activation of Mitogen-activated Protein Kinase*

Treatment of L929 cells with tumor necrosis factor α (TNFα) activates a programmed cell death pathway resulting in apoptosis. We investigated the intracellular signaling pathways activated in L929 cells by TNFα. TNFα robustly activates Jun kinase (JNK), a member of the mitogen-activated protein kinase (MAPK) family. In addition, p42MAPK is activated, but a 10-fold greater concentration of TNFα was required for substantial MAPK activation than was needed for maximal JNK stimulation. Simultaneous treatment of L929 cells with fibroblast growth factor (FGF-2) significantly reduced the apoptotic response to TNFα. FGF-2 substantially activated the Raf/MEK/MAPK (where MEK is mitogen-activated protein kinase kinase) pathway but did not affect TNFα activation of JNK. These results indicate that although JNK may play an important role in transmitting the TNFα signal from the cell surface to the nucleus, activation of the JNK pathway is not sufficient to induce apoptosis. Expression of dominant-negative Asn-17 Ras in L929 cells diminished the FGF-2 stimulation of p42MAPK and eliminated the protective effect of FGF-2. Asn-17 Ras expression did not affect JNK activity and had no effect on TNFα activation of JNK. Pharmacological inhibition of MEK-1 activity by incubation of cells with the compound PD 098059 blocked p42MAPK activation and FGF-2 protection against apoptosis. Interestingly, activated Val-12 Ras expression substantially enhanced TNFα-mediated apoptosis in L929 cells, but Val-12 Ras did not constitutively activate MAPK in L929 cells and FGF-2 partially protected Val-12 Ras-expressing cells from TNFα-mediated apoptosis. Our data indicate that activation of the MAPK pathway mediates an FGF-2 protective effect against apoptosis and highlights the important role that integration of multiple intracellular signaling pathways plays in the regulation of cell growth and death.

[1]  E. Gelfand,et al.  Selective Activation of c-Jun Kinase Mitogen-activated Protein Kinase by CD40 on Human B Cells (*) , 1995, The Journal of Biological Chemistry.

[2]  Michael E. Greenberg,et al.  Opposing Effects of ERK and JNK-p38 MAP Kinases on Apoptosis , 1995, Science.

[3]  Philip R. Cohen,et al.  PD 098059 Is a Specific Inhibitor of the Activation of Mitogen-activated Protein Kinase Kinase in Vitro and in Vivo(*) , 1995, The Journal of Biological Chemistry.

[4]  D. Cortez,et al.  Structural and signaling requirements for BCR-ABL-mediated transformation and inhibition of apoptosis , 1995, Molecular and cellular biology.

[5]  O. Witte,et al.  Alternative signals to RAS for hematopoietic transformation by the BCR-ABL oncogene , 1995, Cell.

[6]  G. Cooper,et al.  Requirement for phosphatidylinositol-3 kinase in the prevention of apoptosis by nerve growth factor. , 1995, Science.

[7]  L. Mahadevan,et al.  Parallel signal processing among mammalian MAPKs. , 1995, Trends in biochemical sciences.

[8]  K. Arai,et al.  Suppression of apoptotic death in hematopoietic cells by signalling through the IL‐3/GM‐CSF receptors. , 1995, The EMBO journal.

[9]  H. K. Sluss,et al.  Signal transduction by tumor necrosis factor mediated by JNK protein kinases , 1994, Molecular and cellular biology.

[10]  L. Greene,et al.  Proliferative inhibition by dominant‐negative Ras rescues naive and neuronally differentiated PC12 cells from apoptotic death. , 1994, The EMBO journal.

[11]  K. Alitalo,et al.  c‐Myc induces cellular susceptibility to the cytotoxic action of TNF‐alpha. , 1994, The EMBO journal.

[12]  M. Karin,et al.  c-Jun N-terminal phosphorylation correlates with activation of the JNK subgroup but not the ERK subgroup of mitogen-activated protein kinases , 1994, Molecular and cellular biology.

[13]  A. Porter,et al.  Nuclear c-Myc plays an important role in the cytotoxicity of tumor necrosis factor alpha in tumor cells , 1994, Molecular and cellular biology.

[14]  T. McDonnell,et al.  Differential sensitivity of normal and Ha-ras-transformed C3H mouse embryo fibroblasts to tumor necrosis factor: induction of bcl-2, c-myc, and manganese superoxide dismutase in resistant cells. , 1994, Oncogene.

[15]  G. Evan,et al.  c‐Myc‐induced apoptosis in fibroblasts is inhibited by specific cytokines. , 1994, The EMBO journal.

[16]  D. Green,et al.  Activation-induced apoptosis in lymphocytes. , 1994, Current opinion in immunology.

[17]  J. Woodgett,et al.  The stress-activated protein kinase subfamily of c-Jun kinases , 1994, Nature.

[18]  I. Behrmann,et al.  Regulation of apoptosis in the immune system. , 1994, Current opinion in immunology.

[19]  Terry Farrah,et al.  The TNF receptor superfamily of cellular and viral proteins: Activation, costimulation, and death , 1994, Cell.

[20]  M. Karin,et al.  Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. , 1993, Genes & development.

[21]  M. McMahon,et al.  Conditional transformation of cells and rapid activation of the mitogen-activated protein kinase cascade by an estradiol-dependent human raf-1 protein kinase , 1993, Molecular and cellular biology.

[22]  L. Tartaglia,et al.  A novel domain within the 55 kd TNF receptor signals cell death , 1993, Cell.

[23]  Wei Li,et al.  Tumor necrosis factor-induced activation and increased tyrosine phosphorylation of mitogen-activated protein (MAP) kinase in human fibroblasts. , 1993, The Journal of biological chemistry.

[24]  R. Davis,et al.  The mitogen-activated protein kinase signal transduction pathway. , 1993, The Journal of biological chemistry.

[25]  S. Nagata,et al.  A novel protein domain required for apoptosis. Mutational analysis of human Fas antigen. , 1993, The Journal of biological chemistry.

[26]  R. Davis,et al.  Transactivation of gene expression by Myc is inhibited by mutation at the phosphorylation sites Thr-58 and Ser-62. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[27]  W. Fiers,et al.  Tumor necrosis factor stimulates multiple serine/threonine protein kinases in Swiss 3T3 and L929 cells. Implication of casein kinase-2 and extracellular signal-regulated kinases in the tumor necrosis factor signal transduction pathway. , 1992, The Journal of biological chemistry.

[28]  M. Karin,et al.  The mammalian ultraviolet response is triggered by activation of src tyrosine kinases , 1992, Cell.

[29]  C. Crews,et al.  The primary structure of MEK, a protein kinase that phosphorylates the ERK gene product. , 1992, Science.

[30]  G. Evan,et al.  Cooperative interaction between c-myc and bcl-2 proto-oncogenes , 1992, Nature.

[31]  C. Crews,et al.  Purification of a murine protein-tyrosine/threonine kinase that phosphorylates and activates the Erk-1 gene product: relationship to the fission yeast byr1 gene product. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[32]  N. Qian,et al.  Mitogen-activated protein kinase activation resulting from selective oncogene expression in NIH 3T3 and rat 1a cells. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Jonathan A. Cooper,et al.  Purification and characterization of mitogen-activated protein kinase activator(s) from epidermal growth factor-stimulated A431 cells. , 1992, The Journal of biological chemistry.

[34]  D. Green,et al.  Role for c-myc in activation-induced apoptotic cell death in T cell hybridomas. , 1992, Science.

[35]  N. Fan,et al.  The p70 tumor necrosis factor receptor mediates cytotoxicity , 1992, Cell.

[36]  Philip R. Cohen,et al.  MAP kinase activator from insulin‐stimulated skeletal muscle is a protein threonine/tyrosine kinase. , 1992, The EMBO journal.

[37]  Gerard I. Evan,et al.  Induction of apoptosis in fibroblasts by c-myc protein , 1992, Cell.

[38]  J. Simpson.  Britain's Chernobyl? , 1992, Nature.

[39]  R. Davis,et al.  A phosphorylation site located in the NH2-terminal domain of c-Myc increases transactivation of gene expression. , 1991, The Journal of biological chemistry.

[40]  James R. Woodgett,et al.  Phosphorylation of c-jun mediated by MAP kinases , 1991, Nature.

[41]  H. Kolb,et al.  Analysis of TNF alpha-induced DNA strand breaks at the single cell level. , 1991, The American journal of pathology.

[42]  E. Chen,et al.  Cloning and expression of cDNAs for two distinct murine tumor necrosis factor receptors demonstrate one receptor is species specific. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[43]  N. Kyprianou,et al.  Activation of programmed cell death by recombinant human tumor necrosis factor plus topoisomerase II-targeted drugs in L929 tumor cells. , 1991, Journal of the National Cancer Institute.

[44]  Y. Hannun,et al.  Identification of sphingomyelin turnover as an effector mechanism for the action of tumor necrosis factor alpha and gamma-interferon. Specific role in cell differentiation. , 1991, The Journal of biological chemistry.

[45]  J. Larrick,et al.  Cytotoxic mechanism of tumor necrosis factor‐α , 1990, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[46]  R. Heller,et al.  Complementary DNA cloning of a receptor for tumor necrosis factor and demonstration of a shed form of the receptor. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[47]  C. Smith,et al.  A receptor for tumor necrosis factor defines an unusual family of cellular and viral proteins. , 1990, Science.

[48]  G. Wong,et al.  Molecular cloning and expression of a receptor for human tumor necrosis factor , 1990, Cell.

[49]  H. Tabuchi,et al.  Molecular cloning and expression of the human 55 kd tumor necrosis factor receptor , 1990, Cell.

[50]  G. Cooper,et al.  Inhibition of NIH 3T3 cell proliferation by a mutant ras protein with preferential affinity for GDP , 1988, Molecular and cellular biology.

[51]  Torti,et al.  Tumor necrosis factor inhibits human myogenesis in vitro , 1988, Molecular and cellular biology.

[52]  J. D. Albert,et al.  Shock and tissue injury induced by recombinant human cachectin. , 1986, Science.

[53]  G. Trinchieri,et al.  Tumor necrosis factor and lymphotoxin induce differentiation of human myeloid cell lines in synergy with immune interferon , 1986, The Journal of experimental medicine.

[54]  J. Vilček,et al.  Fibroblast growth enhancing activity of tumor necrosis factor and its relationship to other polypeptide growth factors , 1986, The Journal of experimental medicine.

[55]  P. Cuatrecasas,et al.  Cellular receptor for 125I-labeled tumor necrosis factor: specific binding, affinity labeling, and relationship to sensitivity. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[56]  Bruce Beutler,et al.  A macrophage factor inhibits adipocyte gene expression: an in vitro model of cachexia. , 1985, Science.

[57]  Eugenio Santos,et al.  A point mutation is responsible for the acquisition of transforming properties by the T24 human bladder carcinoma oncogene , 1982, Nature.

[58]  Cori Bargmann,et al.  Mechanism of activation of a human oncogene , 1982, Nature.

[59]  E. Scolnick,et al.  Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family , 1982, Journal of virology.

[60]  A. Wyllie,et al.  Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics , 1972, British Journal of Cancer.

[61]  N. Finter.  Dye Uptake Methods for Assessing Viral Cytopathogenicity and their Application to Interferon Assays , 1969 .

[62]  G. Johnson,et al.  Measuring activation of kinases in mitogen-activated protein kinase regulatory network. , 1994, Methods in enzymology.

[63]  L. Tartaglia,et al.  Two TNF receptors. , 1992, Immunology today.

[64]  R. Oppenheim.  Cell death during development of the nervous system. , 1991, Annual review of neuroscience.