Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2000 Nov 15;352(Pt 1):135–143.

Lysophosphatidic acid prevents apoptosis in fibroblasts via G(i)-protein-mediated activation of mitogen-activated protein kinase.

X Fang 1, S Yu 1, R LaPushin 1, Y Lu 1, T Furui 1, L Z Penn 1, D Stokoe 1, J R Erickson 1, R C Bast Jr 1, G B Mills 1
PMCID: PMC1221440  PMID: 11062066

Abstract

Lysophosphatidic acid (LPA) is a naturally occurring phospholipid with multiple biological functions. In the present study, we demonstrate that, besides its mitogenic activity, LPA is a potent survival factor, preventing serum-deprivation-induced apoptosis in fibroblasts and other cell types. Both the proliferative effect and survival activity of LPA are sensitive to the action of pertussis toxin (PTX), indicating that both processes are mediated by G(i) protein(s). We therefore focused on the role of G(i)-protein-mediated signalling events in the promotion of cell survival by LPA. In addition to activation of mitogen-activated protein kinase (MAPK), LPA stimulates a modest PTX-sensitive phosphorylation/activation of the serine/threonine kinase Akt, a survival mediator downstream of phosphoinositide 3-kinase (PI3K). Inhibition of PI3K with LY 294002 or wortmannin resulted in a marked inhibition of LPA-induced DNA synthesis, and yet the survival activity of LPA decreased by only 20-30%, suggesting a limited input of the PI3K-Akt cascade in LPA-induced cell survival. In contrast, inhibition of MAPK activation by the MEK-1 inhibitor, PD 98059, blocked both the proliferative and survival effects of LPA. These results indicate that LPA promotes cell survival largely via G(i)-protein-mediated activation of ERK1/ERK2, or other PD 98059-sensitive member(s) of the MAPK family.

Full Text

The Full Text of this article is available as a PDF (366.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andjelković M., Suidan H. S., Meier R., Frech M., Alessi D. R., Hemmings B. A. Nerve growth factor promotes activation of the alpha, beta and gamma isoforms of protein kinase B in PC12 pheochromocytoma cells. Eur J Biochem. 1998 Jan 15;251(1-2):195–200. doi: 10.1046/j.1432-1327.1998.2510195.x. [DOI] [PubMed] [Google Scholar]
  2. Bonni A., Brunet A., West A. E., Datta S. R., Takasu M. A., Greenberg M. E. Cell survival promoted by the Ras-MAPK signaling pathway by transcription-dependent and -independent mechanisms. Science. 1999 Nov 12;286(5443):1358–1362. doi: 10.1126/science.286.5443.1358. [DOI] [PubMed] [Google Scholar]
  3. Brunet A., Bonni A., Zigmond M. J., Lin M. Z., Juo P., Hu L. S., Anderson M. J., Arden K. C., Blenis J., Greenberg M. E. Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell. 1999 Mar 19;96(6):857–868. doi: 10.1016/s0092-8674(00)80595-4. [DOI] [PubMed] [Google Scholar]
  4. Burgering B. M., Coffer P. J. Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction. Nature. 1995 Aug 17;376(6541):599–602. doi: 10.1038/376599a0. [DOI] [PubMed] [Google Scholar]
  5. Cardone M. H., Roy N., Stennicke H. R., Salvesen G. S., Franke T. F., Stanbridge E., Frisch S., Reed J. C. Regulation of cell death protease caspase-9 by phosphorylation. Science. 1998 Nov 13;282(5392):1318–1321. doi: 10.1126/science.282.5392.1318. [DOI] [PubMed] [Google Scholar]
  6. Chou C. C., Yung B. Y. Antiapoptotic effect of ras in the apoptosis induced by serum deprivation and exposure to actinomycin D. Naunyn Schmiedebergs Arch Pharmacol. 1997 Feb;355(2):177–182. doi: 10.1007/pl00004929. [DOI] [PubMed] [Google Scholar]
  7. Craddock B. L., Orchiston E. A., Hinton H. J., Welham M. J. Dissociation of apoptosis from proliferation, protein kinase B activation, and BAD phosphorylation in interleukin-3-mediated phosphoinositide 3-kinase signaling. J Biol Chem. 1999 Apr 9;274(15):10633–10640. doi: 10.1074/jbc.274.15.10633. [DOI] [PubMed] [Google Scholar]
  8. Cuvillier O., Pirianov G., Kleuser B., Vanek P. G., Coso O. A., Gutkind S., Spiegel S. Suppression of ceramide-mediated programmed cell death by sphingosine-1-phosphate. Nature. 1996 Jun 27;381(6585):800–803. doi: 10.1038/381800a0. [DOI] [PubMed] [Google Scholar]
  9. Datta S. R., Dudek H., Tao X., Masters S., Fu H., Gotoh Y., Greenberg M. E. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell. 1997 Oct 17;91(2):231–241. doi: 10.1016/s0092-8674(00)80405-5. [DOI] [PubMed] [Google Scholar]
  10. Dent P., Reardon D. B., Park J. S., Bowers G., Logsdon C., Valerie K., Schmidt-Ullrich R. Radiation-induced release of transforming growth factor alpha activates the epidermal growth factor receptor and mitogen-activated protein kinase pathway in carcinoma cells, leading to increased proliferation and protection from radiation-induced cell death. Mol Biol Cell. 1999 Aug;10(8):2493–2506. doi: 10.1091/mbc.10.8.2493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dudley D. T., Pang L., Decker S. J., Bridges A. J., Saltiel A. R. A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7686–7689. doi: 10.1073/pnas.92.17.7686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eichholtz T., Jalink K., Fahrenfort I., Moolenaar W. H. The bioactive phospholipid lysophosphatidic acid is released from activated platelets. Biochem J. 1993 May 1;291(Pt 3):677–680. doi: 10.1042/bj2910677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Evan G. I., Wyllie A. H., Gilbert C. S., Littlewood T. D., Land H., Brooks M., Waters C. M., Penn L. Z., Hancock D. C. Induction of apoptosis in fibroblasts by c-myc protein. Cell. 1992 Apr 3;69(1):119–128. doi: 10.1016/0092-8674(92)90123-t. [DOI] [PubMed] [Google Scholar]
  14. Fang X., Yu S., Eder A., Mao M., Bast R. C., Jr, Boyd D., Mills G. B. Regulation of BAD phosphorylation at serine 112 by the Ras-mitogen-activated protein kinase pathway. Oncogene. 1999 Nov 18;18(48):6635–6640. doi: 10.1038/sj.onc.1203076. [DOI] [PubMed] [Google Scholar]
  15. Franke T. F., Yang S. I., Chan T. O., Datta K., Kazlauskas A., Morrison D. K., Kaplan D. R., Tsichlis P. N. The protein kinase encoded by the Akt proto-oncogene is a target of the PDGF-activated phosphatidylinositol 3-kinase. Cell. 1995 Jun 2;81(5):727–736. doi: 10.1016/0092-8674(95)90534-0. [DOI] [PubMed] [Google Scholar]
  16. Frankel A., Mills G. B. Peptide and lipid growth factors decrease cis-diamminedichloroplatinum-induced cell death in human ovarian cancer cells. Clin Cancer Res. 1996 Aug;2(8):1307–1313. [PubMed] [Google Scholar]
  17. Gaits F., Fourcade O., Le Balle F., Gueguen G., Gaigé B., Gassama-Diagne A., Fauvel J., Salles J. P., Mauco G., Simon M. F. Lysophosphatidic acid as a phospholipid mediator: pathways of synthesis. FEBS Lett. 1997 Jun 23;410(1):54–58. doi: 10.1016/s0014-5793(97)00411-0. [DOI] [PubMed] [Google Scholar]
  18. Gerrard J. M., Kindom S. E., Peterson D. A., Peller J., Krantz K. E., White J. G. Lysophosphatidic acids. Influence on platelet aggregation and intracellular calcium flux. Am J Pathol. 1979 Aug;96(2):423–438. [PMC free article] [PubMed] [Google Scholar]
  19. Goetzl E. J., An S. A subfamily of G protein-coupled cellular receptors for lysophospholipids and lysosphingolipids. Adv Exp Med Biol. 1999;469:259–264. doi: 10.1007/978-1-4615-4793-8_38. [DOI] [PubMed] [Google Scholar]
  20. Goetzl E. J., Kong Y., Mei B. Lysophosphatidic acid and sphingosine 1-phosphate protection of T cells from apoptosis in association with suppression of Bax. J Immunol. 1999 Feb 15;162(4):2049–2056. [PubMed] [Google Scholar]
  21. Goruppi S., Ruaro E., Schneider C. Gas6, the ligand of Axl tyrosine kinase receptor, has mitogenic and survival activities for serum starved NIH3T3 fibroblasts. Oncogene. 1996 Feb 1;12(3):471–480. [PubMed] [Google Scholar]
  22. Gupta K., Kshirsagar S., Li W., Gui L., Ramakrishnan S., Gupta P., Law P. Y., Hebbel R. P. VEGF prevents apoptosis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling. Exp Cell Res. 1999 Mar 15;247(2):495–504. doi: 10.1006/excr.1998.4359. [DOI] [PubMed] [Google Scholar]
  23. Hetman M., Kanning K., Cavanaugh J. E., Xia Z. Neuroprotection by brain-derived neurotrophic factor is mediated by extracellular signal-regulated kinase and phosphatidylinositol 3-kinase. J Biol Chem. 1999 Aug 6;274(32):22569–22580. doi: 10.1074/jbc.274.32.22569. [DOI] [PubMed] [Google Scholar]
  24. Holmström T. H., Tran S. E., Johnson V. L., Ahn N. G., Chow S. C., Eriksson J. E. Inhibition of mitogen-activated kinase signaling sensitizes HeLa cells to Fas receptor-mediated apoptosis. Mol Cell Biol. 1999 Sep;19(9):5991–6002. doi: 10.1128/mcb.19.9.5991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Holtsberg F. W., Steiner M. R., Bruce-Keller A. J., Keller J. N., Mattson M. P., Moyers J. C., Steiner S. M. Lysophosphatidic acid and apoptosis of nerve growth factor-differentiated PC12 cells. J Neurosci Res. 1998 Sep 15;53(6):685–696. doi: 10.1002/(SICI)1097-4547(19980915)53:6<685::AID-JNR7>3.0.CO;2-1. [DOI] [PubMed] [Google Scholar]
  26. Holtsberg F. W., Steiner M. R., Keller J. N., Mark R. J., Mattson M. P., Steiner S. M. Lysophosphatidic acid induces necrosis and apoptosis in hippocampal neurons. J Neurochem. 1998 Jan;70(1):66–76. doi: 10.1046/j.1471-4159.1998.70010066.x. [DOI] [PubMed] [Google Scholar]
  27. Howe L. R., Marshall C. J. Lysophosphatidic acid stimulates mitogen-activated protein kinase activation via a G-protein-coupled pathway requiring p21ras and p74raf-1. J Biol Chem. 1993 Oct 5;268(28):20717–20720. [PubMed] [Google Scholar]
  28. Imamura F., Horai T., Mukai M., Shinkai K., Sawada M., Akedo H. Induction of in vitro tumor cell invasion of cellular monolayers by lysophosphatidic acid or phospholipase D. Biochem Biophys Res Commun. 1993 Jun 15;193(2):497–503. doi: 10.1006/bbrc.1993.1651. [DOI] [PubMed] [Google Scholar]
  29. Kamakura S., Moriguchi T., Nishida E. Activation of the protein kinase ERK5/BMK1 by receptor tyrosine kinases. Identification and characterization of a signaling pathway to the nucleus. J Biol Chem. 1999 Sep 10;274(37):26563–26571. doi: 10.1074/jbc.274.37.26563. [DOI] [PubMed] [Google Scholar]
  30. Koh J. S., Lieberthal W., Heydrick S., Levine J. S. Lysophosphatidic acid is a major serum noncytokine survival factor for murine macrophages which acts via the phosphatidylinositol 3-kinase signaling pathway. J Clin Invest. 1998 Aug 15;102(4):716–727. doi: 10.1172/JCI1002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kranenburg O., Verlaan I., Hordijk P. L., Moolenaar W. H. Gi-mediated activation of the Ras/MAP kinase pathway involves a 100 kDa tyrosine-phosphorylated Grb2 SH3 binding protein, but not Src nor Shc. EMBO J. 1997 Jun 2;16(11):3097–3105. doi: 10.1093/emboj/16.11.3097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kulik G., Klippel A., Weber M. J. Antiapoptotic signalling by the insulin-like growth factor I receptor, phosphatidylinositol 3-kinase, and Akt. Mol Cell Biol. 1997 Mar;17(3):1595–1606. doi: 10.1128/mcb.17.3.1595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kulik G., Weber M. J. Akt-dependent and -independent survival signaling pathways utilized by insulin-like growth factor I. Mol Cell Biol. 1998 Nov;18(11):6711–6718. doi: 10.1128/mcb.18.11.6711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Levine J. S., Koh J. S., Triaca V., Lieberthal W. Lysophosphatidic acid: a novel growth and survival factor for renal proximal tubular cells. Am J Physiol. 1997 Oct;273(4 Pt 2):F575–F585. doi: 10.1152/ajprenal.1997.273.4.F575. [DOI] [PubMed] [Google Scholar]
  35. Moolenaar W. H., Kruijer W., Tilly B. C., Verlaan I., Bierman A. J., de Laat S. W. Growth factor-like action of phosphatidic acid. Nature. 1986 Sep 11;323(6084):171–173. doi: 10.1038/323171a0. [DOI] [PubMed] [Google Scholar]
  36. Moolenaar W. H. Lysophosphatidic acid, a multifunctional phospholipid messenger. J Biol Chem. 1995 Jun 2;270(22):12949–12952. doi: 10.1074/jbc.270.22.12949. [DOI] [PubMed] [Google Scholar]
  37. Persons D. L., Yazlovitskaya E. M., Cui W., Pelling J. C. Cisplatin-induced activation of mitogen-activated protein kinases in ovarian carcinoma cells: inhibition of extracellular signal-regulated kinase activity increases sensitivity to cisplatin. Clin Cancer Res. 1999 May;5(5):1007–1014. [PubMed] [Google Scholar]
  38. Ridley A. J., Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 1992 Aug 7;70(3):389–399. doi: 10.1016/0092-8674(92)90163-7. [DOI] [PubMed] [Google Scholar]
  39. Scheid M. P., Duronio V. Dissociation of cytokine-induced phosphorylation of Bad and activation of PKB/akt: involvement of MEK upstream of Bad phosphorylation. Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7439–7444. doi: 10.1073/pnas.95.13.7439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Scheid M. P., Schubert K. M., Duronio V. Regulation of bad phosphorylation and association with Bcl-x(L) by the MAPK/Erk kinase. J Biol Chem. 1999 Oct 22;274(43):31108–31113. doi: 10.1074/jbc.274.43.31108. [DOI] [PubMed] [Google Scholar]
  41. Seufferlein T., Rozengurt E. Lysophosphatidic acid stimulates tyrosine phosphorylation of focal adhesion kinase, paxillin, and p130. Signaling pathways and cross-talk with platelet-derived growth factor. J Biol Chem. 1994 Mar 25;269(12):9345–9351. [PubMed] [Google Scholar]
  42. Toews M. L., Ustinova E. E., Schultz H. D. Lysophosphatidic acid enhances contractility of isolated airway smooth muscle. J Appl Physiol (1985) 1997 Oct;83(4):1216–1222. doi: 10.1152/jappl.1997.83.4.1216. [DOI] [PubMed] [Google Scholar]
  43. Vanhaesebroeck B., Waterfield M. D. Signaling by distinct classes of phosphoinositide 3-kinases. Exp Cell Res. 1999 Nov 25;253(1):239–254. doi: 10.1006/excr.1999.4701. [DOI] [PubMed] [Google Scholar]
  44. Weiner J. A., Chun J. Schwann cell survival mediated by the signaling phospholipid lysophosphatidic acid. Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):5233–5238. doi: 10.1073/pnas.96.9.5233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Xia Z., Dickens M., Raingeaud J., Davis R. J., Greenberg M. E. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science. 1995 Nov 24;270(5240):1326–1331. doi: 10.1126/science.270.5240.1326. [DOI] [PubMed] [Google Scholar]
  46. Xu Y., Gaudette D. C., Boynton J. D., Frankel A., Fang X. J., Sharma A., Hurteau J., Casey G., Goodbody A., Mellors A. Characterization of an ovarian cancer activating factor in ascites from ovarian cancer patients. Clin Cancer Res. 1995 Oct;1(10):1223–1232. [PubMed] [Google Scholar]
  47. Xu Y., Shen Z., Wiper D. W., Wu M., Morton R. E., Elson P., Kennedy A. W., Belinson J., Markman M., Casey G. Lysophosphatidic acid as a potential biomarker for ovarian and other gynecologic cancers. JAMA. 1998 Aug 26;280(8):719–723. doi: 10.1001/jama.280.8.719. [DOI] [PubMed] [Google Scholar]
  48. van Corven E. J., Groenink A., Jalink K., Eichholtz T., Moolenaar W. H. Lysophosphatidate-induced cell proliferation: identification and dissection of signaling pathways mediated by G proteins. Cell. 1989 Oct 6;59(1):45–54. doi: 10.1016/0092-8674(89)90868-4. [DOI] [PubMed] [Google Scholar]
  49. van der Bend R. L., de Widt J., van Corven E. J., Moolenaar W. H., van Blitterswijk W. J. The biologically active phospholipid, lysophosphatidic acid, induces phosphatidylcholine breakdown in fibroblasts via activation of phospholipase D. Comparison with the response to endothelin. Biochem J. 1992 Jul 1;285(Pt 1):235–240. doi: 10.1042/bj2850235. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES