Archives of Pharmacal Research

, Volume 31, Issue 5, pp 628–633 | Cite as

Dioeloyl phosphatidic acid induces morphological changes through an endogenous LPA receptor in C6 glioma cells

  • Young-Ja Chang
  • Yu-Lee Kim
  • Ji-Yeong Jo
  • Kim Kyeok
  • Hyo-Lim Kim
  • Dong-Soon Im
Articles Drug Efficacy and Safety
First Online:
Received:
  • 108 Downloads

Abstract

Previously, we suggested that dioleoyl phosphatidic acid (PA) and lysophosphatidic acid (LPA) increased [Ca2+]i through endogenous LPA receptors coupled to pertussis toxin-sensitive G proteins in rat C6 glioma cells. In the present report, we investigated morphological changes and cytotoxicity induced by PA and LPA in C6 glioma cells. Isoproterenol treatment led to changes in the cell morphology of rat C6 glioma cells, which were reverted by the addition of PA and LPA. PA-and LPA-induced morphological reversions were inhibited by treatment with Ki16425, an LPA1/LPA3 receptor antagonist. VPC32183, another LPA1/LPA3 receptor antagonist with a different structure, only inhibited PA-induced morphological reversion but not LPA-induced reversion. However, the reversions were not inhibited by treatment with pertussis toxin, a specific inhibitor of Gi/o proteins. In addition, cytotoxicity was only induced by LPA but not by PA in C6 glioma cells. Our results suggest that PA may act as a partial agonist at endogenous LPA receptors, which are sensitive to Ki16425 and coupled to PTX-insensitive G proteins, to evoke morphological changes in C6 glioma cells.

Key words

Phosphatidic acid Lysophosphatidic acid G-Protein-coupled receptor Cell shape LPA1 LPA3 

Abbreviations

PA

dioleoyl phosphatidic acid

LPA

lysophosphatidic acid

VPC32183

(S)-Phosphoric acid mono-{2-octadec-9-enoylamino-3-[4-(pyridine-2-ylmethoxy)-phenyl]-propyl} ester

Ki16425

3-(4-[4-([1-(2-Chlorophenyl)ethoxy]carbonylamino)-3-methyl-5-isoxazolyl]benzylthio) propanoic acid

PTX

pertussis toxin

GPCR

G-protein-coupled receptor

LPA1

lysophosphatidic acid receptor type 1 (EDG2)

LPA2

lysophosphatidic acid receptor type 2 (EDG4)

LPA3

lysophosphatidic acid receptor 3 (EDG7)

LPA4

lysophosphatidic acid receptor type 4 (GPR23)

LPA5

lysophosphatidic acid receptor 5 (GPR92)

PLD

phospholipase D

PLA2

phospholipase A2

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References

  1. An, S., Bleu, T., Hallmark, O. G., and Goetzl, E. J., Characterization of a novel subtype of human G protein-coupled receptor for lysophosphatidic acid. J. Biol. Chem., 273, 7906–7910 (1998a).PubMedCrossRefGoogle Scholar
  2. An, S., Bleu, T., Zheng, Y., and Goetzl, E. J., Recombinant human G protein-coupled lysophosphatidic acid receptors mediate intracellular calcium mobilization. Mol. Pharmacol., 54, 881–888 (1998b).PubMedGoogle Scholar
  3. Bashir, N., Kuhen, K., and Taub, M., Phospholipids regulate growth and function of MDCK cells in hormonally defined serum free medium. In Vitro Cell Dev. Biol., 28A, 663–668 (1992).PubMedCrossRefGoogle Scholar
  4. Buckland, A. G. and Wilton, D. C., Anionic phospholipids, interfacial binding and the regulation of cell functions. Biochim. Biophys Acta., 1483, 199–216 (2000).PubMedGoogle Scholar
  5. Chang, Y. J., Lee, Y. K., Lee, E. H., Park, J. J., Chung, S. K., and Im, D. S., Structure-activity relationships of dimethylsphingosine (DMS) derivatives and their effects on intracellular pH and Ca2+ in the U937 monocyte cell line. Arch. Pharm. Res., 29, 657–665 (2006).PubMedGoogle Scholar
  6. Chang, Y. J., Kim, H. L., Sacket, S. J., Kim, K., Han, M., Jo, J. Y., and Im, D. S., Calcium signaling of dioleoyl phosphatidic acid via endogenous LPA receptors: A study using HCT116 and HT29 human colon cancer cell lines. J. Appl. Pharmacol., 15, 150–155 (2007a).CrossRefGoogle Scholar
  7. Chang, Y. J., Kim, Y. L., Lee, Y. K., Sacket, S. J., Kim, K., Kim, H. L., Han, M., Bae, Y. S., Okajima, F., and Im, D. S., Dioleoyl phosphatidic acid increases intracellular Ca2+ through endogenous LPA receptors in C6 glioma and L2071 fibroblasts. Prostaglandins Other Lipid Mediat., 83, 268–276 (2007b).PubMedCrossRefGoogle Scholar
  8. Durgam, G. G., Tsukahara, R., Makarova, N., Walker, M. D., Fujiwara, Y., Pigg, K. R., Baker, D. L., Sardar, V. M., Parrill, A. L., Tigyi, G., and Miller, D. D., Synthesis and pharmacological evaluation of second-generation phosphatidic acid derivatives as lysophosphatidic acid receptor ligands. Bioorg. Med. Chem. Lett., 16, 633–640 (2006).PubMedCrossRefGoogle Scholar
  9. English, D., Cui, Y., and Siddiqui, R. A., Messenger functions of phosphatidic acid. Chem Phys Lipids 80, 117–132 (1996).PubMedCrossRefGoogle Scholar
  10. English, D., Martin, M., Harvey, K. A., Akard, L. P., Allen, R., Widlanski, T. S., Garcia, J. G., and Siddiqui, R. A., Characterization and purification of neutrophil ecto-phosphatidic acid phosphohydrolase. Biochem. J., 324(Pt 3), 941–950 (1997).PubMedGoogle Scholar
  11. Fang, Y., Vilella-Bach, M., Bachmann, R., Flanigan, A., and Chen, J., Phosphatidic acid-mediated mitogenic activation of mTOR signaling. Science, 294, 1942–1945 (2001).PubMedCrossRefGoogle Scholar
  12. Fischer, D. J., Nusser, N., Virag, T., Yokoyama, K., Wang, D., Baker, D. L., Bautista, D., Parrill, A. L., and Tigyi, G., Short-chain phosphatidates are subtype-selective antagonists of lysophosphatidic acid receptors. Mol. Pharmacol., 60, 776–784 (2001).PubMedGoogle Scholar
  13. Fitzgerald, L. R., Dytko, G. M., Sarau, H. M., Mannan, I. J., Ellis, C., Lane, P. A., Tan, K. B., Murdock, P. R., Wilson, S., Bergsma, D. J., Ames, R. S., Foley, J. J., Campbell, D. A., McMillan, L., Evans, N., Elshourbagy, N. A., Minehart, H., and Tsui, P., Identification of an EDG7 variant, HOFNH30, a G-protein-coupled receptor for lysophosphatidic acid. Biochem. Biophys Res. Commun., 273, 805–810 (2000).PubMedCrossRefGoogle Scholar
  14. Gerrard, J. M., Butler, A. M., Peterson, D. A., and White, J. G., Phosphatidic acid releases calcium from a platelet membrane fraction in vitro. Prostaglandins Med., 1, 387–396 (1978).PubMedCrossRefGoogle Scholar
  15. Ha, K. S. and Exton, J. H., Activation of actin polymerization by phosphatidic acid derived from phosphatidylcholine in IIC9 fibroblasts. J. Cell Biol., 123, 1789–1796 (1993).PubMedCrossRefGoogle Scholar
  16. Hiramatsu, T., Sonoda, H., Takanezawa, Y., Morikawa, R., Ishida, M., Kasahara, K., Sanai, Y., Taguchi, R., Aoki, J., and Arai, H., Biochemical and molecular characterization of two phosphatidic acid-selective phospholipase A1s, mPA-PLA1alpha and mPA-PLA1beta. J. Biol. Chem., 278, 49438–49447 (2003).PubMedCrossRefGoogle Scholar
  17. Hornberger, T. A., Chu, W. K., Mak, Y. W., Hsiung, J. W., Huang, S. A., and Chien, S., The role of phospholipase D and phosphatidic acid in the mechanical activation of mTOR signaling in skeletal muscle. Proc. Natl. Acad. Sci. U.S.A., 103, 4741–4746 (2006).PubMedCrossRefGoogle Scholar
  18. Huang, K. S., Li, S., and Low, M. G., Glycosylphosphatidylinositol-specific phospholipase D. Methods Enzymol., 197, 567–575 (1991).PubMedGoogle Scholar
  19. Ikeda, Y., Kikuchi, M., Toyama, K., Watanabe, K., and Ando, Y., Ionophoretic activities of phospholipids on human platelets. Thromb Haemost., 41, 779–786 (1979).PubMedGoogle Scholar
  20. Imai, A., Ishizuka, Y., Kawai, K., and Nozawa, Y., Evidence for coupling of phosphatidic acid formation and calcium influx in thrombin-activated human platelets. Biochem. Biophys. Res. Commun., 108, 752–759 (1982).PubMedCrossRefGoogle Scholar
  21. Kim, H. S., Lee, E. H., Ko, S. R., Choi, K. J., Park, J. H., and Im, D. S., Effects of ginsenosides Rg3 and Rh2 on the proliferation of prostate cancer cells. Arch. Pharm. Res., 27, 429–435 (2004).PubMedGoogle Scholar
  22. Kim, K., Kim, Y. L., Sacket, S. J., Kim, H. L., Han, M., Park, D. S., Lee, B. K., Lee, W. K., Ha, H. J., and Im, D. S., Sphingosine 1-phosphate (S1P) induces shape change in rat C6 glioma cells through the S1P2 receptor: development of an agonist for S1P receptors. J. Pharm. Pharmacol., 59, 1035–1041 (2007).PubMedCrossRefGoogle Scholar
  23. Knauss, T. C., Jaffer, F. E., and Abboud, H. E., Phosphatidic acid modulates DNA synthesis, phospholipase C, and platelet-derived growth factor mRNAs in cultured mesangial cells. Role of protein kinase C. J. Biol. Chem., 265, 14457–14463 (1990).PubMedGoogle Scholar
  24. Kotarsky, K., Boketoft, A., Bristulf, J., Nilsson, N. E., Norberg, A., Hansson, S., Sillard, R., Owman, C., Leeb-Lundberg, F. L., and Olde, B., Lysophosphatidic Acid Binds to and Activates GPR92, a G Protein-Coupled Receptor Highly Expressed in Gastro-Intestinal Lymphocytes. J. Pharmacol. Exp. Ther., 318, 619–628 (2006).PubMedCrossRefGoogle Scholar
  25. Krabak, M. J. and Hui, S. W., The mitogenic activities of phosphatidate are acyl-chain-length dependent and calcium independent in C3H/10T1/2 cells. Cell Regul., 2, 57–64 (1991).PubMedGoogle Scholar
  26. Lee, C. W., Rivera, R., Gardell, S., Dubin, A. E., and Chun, J., GPR92 as a New G12/13- and Gq-coupled Lysophosphatidic Acid Receptor That Increases cAMP, LPA5. J. Biol. Chem., 281, 23589–23597 (2006).PubMedCrossRefGoogle Scholar
  27. Noguchi, K., Ishii, S., and Shimizu, T., Identification of p2y9/GPR23 as a novel G protein-coupled receptor for Lysophosphatidic acid, structurally distant from the Edg family. J. Biol. Chem., 278, 25600–25606 (2003).PubMedCrossRefGoogle Scholar
  28. Oey, J., Noradrenaline induces morphological alterations in nucleated and enucleated rat C6 glioma cells. Nature, 257, 317–319 (1975).PubMedCrossRefGoogle Scholar
  29. Pearce, B., Jakobson, K., Morrow, C., and Murphy, S., Phosphatidic acid promotes phosphoinositide metabolism and DNA synthesis in cultured cortical astrocytes. Neurochem Int., 24, 165–171 (1994).PubMedCrossRefGoogle Scholar
  30. Ryder, N. S., Talwar, H. S., Reynolds, N. J., Voorhees, J. J., and Fisher, G. J., Phosphatidic acid and phospholipase D both stimulate phosphoinositide turnover in cultured human keratinocytes. Cell Signal., 5, 787–794 (1993).PubMedCrossRefGoogle Scholar
  31. Shain, W., Forman, D. S., Madelian, V., and Turner, J. N., Morphology of astroglial cells is controlled by beta-adrenergic receptors. J. Cell Biol., 105, 2307–2314 (1987).PubMedCrossRefGoogle Scholar
  32. Siegmann, D. W., Stimulation of quiescent 3T3 cells by phosphatidic acid-containing liposomes. Biochem. Biophys. Res. Commun., 145, 228–233 (1987).PubMedCrossRefGoogle Scholar
  33. Sonoda, H., Aoki, J., Hiramatsu, T., Ishida, M., Bandoh, K., Nagai, Y., Taguchi, R., Inoue, K., and Arai, H., A novel phosphatidic acid-selective phospholipase A1 that produces lysophosphatidic acid. J. Biol. Chem., 277, 34254–34263 (2002).PubMedCrossRefGoogle Scholar
  34. Stace, C. L. and Ktistakis, N. T., Phosphatidic acid-and phosphatidylserine-binding proteins. Biochim. Biophys Acta., 1761, 913–926 (2006).PubMedGoogle Scholar
  35. Tas, P. W. and Koschel, K., Thrombin reverts the beta-adrenergic agonist-induced morphological response in rat glioma C6 cells. Exp. Cell Res., 189, 22–27 (1990).PubMedCrossRefGoogle Scholar
  36. Tas, P. W. and Koschel, K., Sphingosine-1-phosphate induces a Ca2+ signal in primary rat astrocytes and a Ca2+ signal and shape changes in C6 rat glioma cells. J. Neurosci Res., 52, 427–434 (1998).PubMedCrossRefGoogle Scholar
  37. Wood, C. A., Padmore, L., and Radda, G. K., The effect of phosphatidic acid on the proliferation of Swiss 3T3 cells. Biochem. Soc. Trans., 21, 369S (1993).PubMedGoogle Scholar
  38. Yanagida, K., Ishii, S., Hamano, F., Noguchi, K., and Shimizu, T., LPA4/p2y9/GPR23 mediates Rho-dependent morphological changes in a rat neuronal cell line. J. Biol. Chem., 282, 5814–5824 (2007).PubMedCrossRefGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2008

Authors and Affiliations

  • Young-Ja Chang
    • 1
  • Yu-Lee Kim
    • 1
  • Ji-Yeong Jo
    • 1
  • Kim Kyeok
    • 1
  • Hyo-Lim Kim
    • 1
  • Dong-Soon Im
    • 1
  1. 1.Laboratory of Pharmacology, College of Pharmacy (BK21 Project) and Longevity Life Science and Technology InstitutesPusan National UniversityBusanRepublic of Korea

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