Protein kinase C mediates basic fibroblast growth factor protection of endothelial cells against radiation-induced apoptosis.

Basic fibroblast growth factor (bFGF) was found to protect bovine aortic endothelial cells against the lethal effects of ionizing radiation by inhibiting the programmed cell death (apoptosis) induced in these cells by radiation exposure. The involvement of the bFGF receptor tyrosine kinase in this function was demonstrated by abrogation of the radioprotective effect of bFGF by a specific inhibitor of the bFGF receptor tyrosine kinase, the tyrphostin AG213. The downstream signaling after stimulation of the bFGF receptor tyrosine kinase in bovine aortic endothelial cells involved translocation of the alpha isotype of cytoplasmic protein kinase C (PKC) into the membrane and its activation within 30 s after bFGF stimulation. The involvement of PKC in the radioprotective effect conferred by bFGF was suggested by the demonstration that nonspecific PKC activation by short-term exposure (30 min) to the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA; 30 ng/ml) mimicked the radioprotective effect of bFGF. Furthermore, treatment of the cells with the PKC inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (20 microM) abrogated the radioprotective effect of bFGF, as was observed after the depletion of cellular PKC by overnight preincubation with high-dose TPA (200 nM). Agarose gel electrophoresis of DNA extracted from irradiated bovine aortic endothelial cells showed that both TPA (30 ng/ml; 30 min) and bFGF (1 ng/ml) inhibited the apoptotic degradation of DNA induced in these cells by radiation exposure (500 cGy). Both the bFGF- and the TPA-mediated inhibition of apoptosis could be reversed by the PKC inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (20 microM). These data demonstrate the involvement of PKC in the inhibition of radiation-induced apoptosis by bFGF and the rescue of endothelial cells from this mode of radiation-induced cell death.

[1]  J. Schwartz,et al.  Basic fibroblast growth factor protects endothelial cells against radiation-induced programmed cell death in vitro and in vivo. , 1994, Cancer research.

[2]  G. Schieven,et al.  Tyrosine phosphorylation is a mandatory proximal step in radiation-induced activation of the protein kinase C signaling pathway in human B-lymphocyte precursors , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[3]  J. Thiery,et al.  Cyclic AMP distinguishes between two functions of acidic FGF in a rat bladder carcinoma cell line , 1993, The Journal of cell biology.

[4]  A. Levitzki.  Tyrphostins: tyrosine kinase blockers as novel antiproliferative agents and dissectors of signal transduction , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  B. Strulovici,et al.  Differential expression of protein kinase C epsilon protein in lung cancer cell lines by ionising radiation. , 1992, British Journal of Cancer.

[6]  Y. Nishizuka.  Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. , 1992, Science.

[7]  Nanxin Li,et al.  Point mutation in FGF receptor eliminates phosphatidylinositol hydrolysis without affecting mitogenesis , 1992, Nature.

[8]  S. Cook,et al.  Epidermal growth factor increases sn-1,2-diacylglycerol levels and activates phospholipase D-catalysed phosphatidylcholine breakdown in Swiss 3T3 cells in the absence of inositol-lipid hydrolysis. , 1992, The Biochemical journal.

[9]  A. Haman,et al.  Role of protein kinase C and the Na+/H+ antiporter in suppression of apoptosis by granulocyte macrophage colony-stimulating factor and interleukin-3. , 1992, The Journal of biological chemistry.

[10]  D. Hallahan,et al.  Inhibition of protein kinases sensitizes human tumor cells to ionizing radiation. , 1992, Radiation research.

[11]  B. Pettmann,et al.  Phosphorylation of the MARCKS Protein (P87), a Major Protein Kinase C Substrate, Is Not an Obligatory Step in the Mitogenic Signaling Pathway of Basic Fibroblast Growth Factor in Rat Oligodendrocytes , 1992, Journal of neurochemistry.

[12]  M. Klagsbrun,et al.  The Fibroblast Growth Factor Family An Overview , 1991 .

[13]  M. Auffredou,et al.  Differential inhibition of interleukin 2‐ and interleukin 4‐mediated human B cell proliferation by ionomycin: A possible regulatory role for apoptosis , 1991, European journal of immunology.

[14]  D. Hallahan,et al.  Tumor necrosis factor gene expression is mediated by protein kinase C following activation by ionizing radiation. , 1991, Cancer research.

[15]  J. Lotem,et al.  Rescue from programmed cell death in leukemic and normal myeloid cells. , 1991, Blood.

[16]  S. Orrenius,et al.  Inhibition of DNA fragmentation in thymocytes and isolated thymocyte nuclei by agents that stimulate protein kinase C. , 1989, The Journal of biological chemistry.

[17]  C. Wenner,et al.  Inhibition of radiation-induced apoptosis in vitro by tumor promoters. , 1988, Biochemical and biophysical research communications.

[18]  E. Rozengurt.  Early signals in the mitogenic response. , 1986, Science.

[19]  D. Stumpo,et al.  Effects of mitogens on ornithine decarboxylase activity and messenger RNA levels in normal and protein kinase C-deficient NIH-3T3 fibroblasts. , 1986, The Journal of biological chemistry.

[20]  E. Krebs,et al.  Fibroblast growth factor treatment of Swiss 3T3 cells activates a subunit S6 kinase that phosphorylates a synthetic peptide substrate. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Y. Nishizuka.  Studies and perspectives of protein kinase C. , 1986, Science.

[22]  K. Kaibuchi,et al.  Possible involvement of protein kinase C and calcium ion in growth factor-induced expression of c-myc oncogene in Swiss 3T3 fibroblasts. , 1986, The Journal of biological chemistry.

[23]  P. Blackshear,et al.  Growth factor-stimulated protein phosphorylation in 3T3-L1 cells. Evidence for protein kinase C-dependent and -independent pathways. , 1985, The Journal of biological chemistry.

[24]  A. Wyllie,et al.  Macrophage recognition of cells undergoing programmed cell death (apoptosis). , 1985, Immunology.

[25]  S. Kawamoto,et al.  Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. , 1984, Biochemistry.

[26]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[27]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[28]  A. Haimovitz-Friedman,et al.  Radiation Damage in Endothelial Cells Autocrine Effects of Fibroblast Growth Factor in Repair of Updated , 2006 .

[29]  D. Moscatelli,et al.  The FGF family of growth factors and oncogenes. , 1992, Advances in cancer research.

[30]  D. Hallahan,et al.  The isoquinoline sulfonamide H7 attenuates radiation-mediated protein kinase C activation and delays the onset of x-ray-induced G2 arrest. , 1992, International journal of radiation oncology, biology, physics.

[31]  A. Wyllie.  The biology of cell death in tumours. , 1985, Anticancer research.

[32]  吉川 潮.  Calcium-activated, phospholipid-dependent protein kinase from rat brain , 1984 .