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Lysophosphatidic Acid Stimulates Activation of Focal Adhesion Kinase and Paxillin and Promotes Cell Motility, via LPA1–3, in Human Pancreatic Cancer

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Abstract

Background

Pancreatic cancer is highly metastatic and with poor prognosis. In previous studies, lysophosphatidic acid (LPA) was shown to be a critical component of ascites which promoted the invasion and metastasis of pancreatic cancer. Two focal adhesion proteins, focal adhesion kinase (FAK) and paxillin, were crucially involved in cell migration, cytoskeleton reorganization, and the dynamics of focal adhesion.

Objectives

This study examined the involvement of LPA1–3 in LPA-induced activation of FAK and paxillin, and in cell motility, in pancreatic cancer PANC-1 cells.

Methods

Reverse transcriptase polymerase chain reaction analysis was used to examine mRNA expression of LPA receptors in PANC-1. Cellular protein expression of FAK and paxillin was analyzed by western blotting. The subcellular location of FAK and paxillin was visualized by immunofluorescence. Cell migration was measured by use of a transwell migration chamber.

Results

Three LPA receptors (LPA1, LPA2, and LPA3) were significantly expressed in PANC-1 cells. Treatment with LPA induced both time and dose-dependent tyrosine phosphorylation of FAK and paxillin. LPA also affected translocation of FAK and paxillin from cytoplasm to focal adhesions at the cell periphery and enhanced cell motility of PANC-1. Pretreatment with 3-(4-(4-((1-(2-chlorophenyl)ethoxy)carbonyl amino)-3-methyl-5-isoxazolyl)benzylsulfanyl)propanoic acid (Ki16425), an antagonist of LPA1 and LPA3, before LPA attenuated the LPA-induced tyrosine phosphorylation and redistribution of FAK and paxillin and abrogated LPA-induced cellular migration activity.

Conclusions

These results suggest LPA induces activation of FAK and paxillin via LPA1–3, which may contribute to the increased cell motility in human pancreatic cancer PANC-1 cells. Thus, an understanding of the regulation by LPA of cell motility in pancreatic cancer could identify novel targets for therapy.

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References

  1. Ryder NM, Guha S, Hines OJ, et al. G protein-coupled receptor signaling in human ductal pancreatic cancer cells: neurotensin responsiveness and mitogenic stimulation. J Cell Physiol. 2001;186:53–64.

    Article  PubMed  CAS  Google Scholar 

  2. Komachi M, Tomura H, Malchinkhuu E, et al. LPA1 receptors mediate stimulation, whereas LPA2 receptors mediate inhibition, of migration of pancreatic cancer cells in response to lysophosphatidic acid and malignant ascites. Carcinogenesis. 2009;30:457–465.

    Article  PubMed  CAS  Google Scholar 

  3. Kim MH, Park JS, Chang HJ, et al. Lysophosphatidic acid promotes cell invasion by up-regulating the urokinase-type plasminogen activator receptor in human gastric cancer cells. J Cell Biochem. 2008;104:1102–1112.

    Article  PubMed  CAS  Google Scholar 

  4. Ramachandran S, Shida D, Nagahashi M, et al. Lysophosphatidic acid stimulates gastric cancer cell proliferation via ERK1-dependent upregulation of sphingosine kinase 1 transcription. FEBS Lett. 2010;584:4077–4082.

    Article  PubMed  CAS  Google Scholar 

  5. Shida D, Fang X, Kordula T, et al. Cross-talk between LPA1 and epidermal growth factor receptors mediates up-regulation of sphingosine kinase 1 to promote gastric cancer cell motility and invasion. Cancer Res. 2008;68:6569–6577.

    Article  PubMed  CAS  Google Scholar 

  6. Zhang R, Wang J, Ma S, et al. Requirement of Osteopontin in the migration and protection against Taxol-induced apoptosis via the ATX-LPA axis in SGC7901 cells. BMC Cell Biol. 2011;12:11.

    Article  PubMed  CAS  Google Scholar 

  7. Zhang H, Bialkowska A, Rusovici R, et al. Lysophosphatidic acid facilitates proliferation of colon cancer cells via induction of Krüppel-like factor 5. J Biol Chem. 2007;282:15541–15549.

    Article  PubMed  CAS  Google Scholar 

  8. Yamada T, Sato K, Komachi M, et al. Lysophosphatidic acid (LPA) in malignant ascites stimulates motility of human pancreatic cancer cells through LPA1. J Biol Chem. 2004;279:6595–6605.

    Article  PubMed  CAS  Google Scholar 

  9. Komachi M, Sato K, Tobo M, et al. Orally active lysophosphatidic acid receptor antagonist attenuates pancreatic cancer invasion and metastasis in vivo. Cancer Sci. 2012;103:1099–1104.

    Article  PubMed  CAS  Google Scholar 

  10. Leve F, Marcondes TG, Bastos LG, et al. Lysophosphatidic acid induces a migratory phenotype through a crosstalk between RhoA-Rock and Src-FAK signalling in colon cancer cells. Eur J Pharmacol. 2011;671:7–17.

    Article  PubMed  CAS  Google Scholar 

  11. Iwanicki MP, Vomastek T, Tilghman RW, et al. FAK, PDZ-RhoGEF and ROCKII cooperate to regulate adhesion movement and trailing-edge retraction in fibroblasts. J Cell Sci. 2008;121:895–905.

    Article  PubMed  CAS  Google Scholar 

  12. Jiang X, Jacamo R, Zhukova E, et al. RNA interference reveals a differential role of FAK and Pyk2 in cell migration, leading edge formation and increase in focal adhesions induced by LPA in intestinal epithelial cells. J Cell Physiol. 2006;207:816–828.

    Article  PubMed  CAS  Google Scholar 

  13. Nakamura K, Yano H, Uchida H, et al. Tyrosine phosphorylation of paxillin alpha is involved in temporospatial regulation of paxillin-containing focal adhesion formation and F-actin organization in motile cells. J Biol Chem. 2000;275:27155–27164.

    PubMed  CAS  Google Scholar 

  14. Sawada K, Morishige K, Tahara M, et al. Lysophosphatidic acid induces focal adhesion assembly through Rho/Rho-associated kinase pathway in human ovarian cancer cells. Gynecol Oncol. 2002;87:252–259.

    Article  PubMed  CAS  Google Scholar 

  15. Hashimoto K, Morishige K, Sawada K, et al. Geranylgeranylacetone inhibits lysophosphatidic acid-induced invasion of human ovarian carcinoma cells in vitro. Cancer. 2005;103:1529–1536.

    Article  PubMed  CAS  Google Scholar 

  16. Arita Y, Ito T, Oono T, et al. Lysophosphatidic acid induced nuclear translocation of nuclear factor-kappaB in Panc-1 cells by mobilizing cytosolic free calcium. World J Gastroenterol. 2008;14:4473–4479.

    Article  PubMed  CAS  Google Scholar 

  17. Stähle M, Veit C, Bachfischer U, et al. Mechanisms in LPA-induced tumor cell migration: critical role of phosphorylated ERK. J Cell Sci. 2003;116:3835–3846.

    Article  PubMed  Google Scholar 

  18. Lange K, Kammerer M, Saupe F, et al. Combined lysophosphatidic acid/platelet-derived growth factor signaling triggers glioma cell migration in a tenascin-C microenvironment. Cancer Res. 2008;68:6942–6952.

    Article  PubMed  CAS  Google Scholar 

  19. Salazar EP, Rozengurt E. Src family kinases are required for integrin-mediated but not for G protein-coupled receptor stimulation of focal adhesion kinase autophosphorylation at Tyr-397. J Biol Chem. 2001;276:17788–17795.

    Article  PubMed  CAS  Google Scholar 

  20. Iwasaki T, Nakata A, Mukai M, et al. Involvement of phosphorylation of Tyr-31 and Tyr-118 of paxillin in MM1 cancer cell migration. Int J Cancer. 2002;97:330–335.

    Article  PubMed  CAS  Google Scholar 

  21. Hetey SE, Lalonde DP, Turner CE. Tyrosine-phosphorylated Hic-5 inhibits epidermal growth factor-induced lamellipodia formation. Exp Cell Res. 2005;311:147–156.

    Article  PubMed  CAS  Google Scholar 

  22. Moolenaar WH. Lysophosphatidic acid, a multifunctional phospholipid messenger. J Biol Chem. 1995;270:12949–12952.

    Article  PubMed  CAS  Google Scholar 

  23. Goetzl EJ, An S. Diversity of cellular receptors and functions for the lysophospholipid growth factors lysophosphatidic acid and sphingosine 1-phosphate. FASEB J. 1998;12:1589–1598.

    PubMed  CAS  Google Scholar 

  24. Sun H, Ren J, Zhu Q, et al. Effects of lysophosphatidic acid on human colon cancer cells and its mechanisms of action. World J Gastroenterol. 2009;15:4547–4555.

    Article  PubMed  CAS  Google Scholar 

  25. Shida D, Kitayama J, Yamaguchi H, et al. Lysophosphatidic acid (LPA) enhances the metastatic potential of human colon carcinoma DLD1 cells through LPA1. Cancer Res. 2003;63:1706–1711.

    PubMed  CAS  Google Scholar 

  26. Gibbs TC, Rubio MV, Zhang Z, et al. Signal transduction responses to lysophosphatidic acid and sphingosine 1-phosphate in human prostate cancer cells. Prostate. 2009;69:1493–1506.

    Article  PubMed  CAS  Google Scholar 

  27. Shin KJ, Kim YL, Lee S, et al. Lysophosphatidic acid signaling through LPA receptor subtype 1 induces colony scattering of gastrointestinal cancer cells. J Cancer Res Clin Oncol. 2009;135:45–52.

    Article  PubMed  CAS  Google Scholar 

  28. Shida D, Kitayama J, Yamaguchi H, et al. Dual mode regulation of migration by lysophosphatidic acid in human gastric cancer cells. Exp Cell Res. 2004;301:168–178.

    Article  PubMed  CAS  Google Scholar 

  29. Ward JD, Dhanasekaran DN. LPA stimulates the phosphorylation of p130Cas via Gαi2 in ovarian cancer cells. Genes Cancer. 2012;3:578–591.

    Article  PubMed  Google Scholar 

  30. Jeong KJ, Park SY, Cho KH, et al. The Rho/ROCK pathway for lysophosphatidic acid-induced proteolytic enzyme expression and ovarian cancer cell invasion. Oncogene. 2012;31:4279–4289.

    Article  PubMed  CAS  Google Scholar 

  31. Li TT, Alemayehu M, Aziziyeh AI, et al. Beta-arrestin/Ral signaling regulates lysophosphatidic acid-mediated migration and invasion of human breast tumor cells. Mol Cancer Res. 2009;7:1064–1077.

    Article  PubMed  CAS  Google Scholar 

  32. Tanabe E, Kitayoshi M, Yoshikawa K, et al. Loss of lysophosphatidic acid receptor-3 suppresses cell migration activity of human sarcoma cells. J Recept Signal Transduct Res. 2012;32:328–334.

    Article  PubMed  CAS  Google Scholar 

  33. Jung ID, Lee J, Lee KB, et al. Activation of p21-activated kinase 1 is required for lysophosphatidic acid-induced focal adhesion kinase phosphorylation and cell motility in human melanoma A2058 cells. Eur J Biochem. 2004;271:1557–1565.

    Article  PubMed  CAS  Google Scholar 

  34. Rusovici R, Ghaleb A, Shim H, et al. Lysophosphatidic acid prevents apoptosis of Caco-2 colon cancer cells via activation of mitogen-activated protein kinase and phosphorylation of Bad. Biochim Biophys Acta. 2007;1770:1194–1203.

    Article  PubMed  CAS  Google Scholar 

  35. Samadi N, Gaetano C, Goping IS, et al. Autotaxin protects MCF-7 breast cancer and MDA-MB-435 melanoma cells against Taxol-induced apoptosis. Oncogene. 2009;28:1028–1039.

    Article  PubMed  CAS  Google Scholar 

  36. Kam Y, Guess C, Estrada L, et al. A novel circular invasion assay mimics in vivo invasive behavior of cancer cell lines and distinguishes single-cell motility in vitro. BMC Cancer. 2008;8:198.

    Article  PubMed  Google Scholar 

  37. Gardner JA, Ha JH, Jayaraman M, et al. The gep proto-oncogene Gα13 mediates lysophosphatidic acid-mediated migration of pancreatic cancer cells. Pancreas. 2013;42:819–828.

    Article  PubMed  CAS  Google Scholar 

  38. Kato K, Yoshikawa K, Tanabe E, et al. Opposite roles of LPA1 and LPA3 on cell motile and invasive activities of pancreatic cancer cells. Tumour Biol. 2012;33:1739–1744.

    Article  PubMed  CAS  Google Scholar 

  39. Yoshikawa K, Tanabe E, Shibata A, et al. Involvement of oncogenic K-ras on cell migration stimulated by lysophosphatidic acid receptor-2 in pancreatic cancer cells. Exp Cell Res. 2013;319:105–112.

    Article  PubMed  CAS  Google Scholar 

  40. Hama K, Aoki J, Fukaya M, et al. Lysophosphatidic acid and autotaxin stimulate cell motility of neoplastic and non-neoplastic cells through LPA1. J Biol Chem. 2004;279:17634–17639.

    Article  PubMed  CAS  Google Scholar 

  41. Hayashi M, Okabe K, Kato K, et al. Differential function of lysophosphatidic acid receptors in cell proliferation and migration of neuroblastoma cells. Cancer Lett. 2012;316:91–96.

    Article  PubMed  CAS  Google Scholar 

  42. Yamashita H, Kitayama J, Shida D, et al. Differential expression of lysophosphatidic acid receptor-2 in intestinal and diffuse type gastric cancer. J Surg Oncol. 2006;93:30–35.

    Article  PubMed  CAS  Google Scholar 

  43. Luttrell LM, Daaka Y, Della Rocca GJ, et al. G protein-coupled receptors mediate two functionally distinct pathways of tyrosine phosphorylation in rat 1a fibroblasts. Shc phosphorylation and receptor endocytosis correlate with activation of Erk kinases. J Biol Chem. 1997;272:31648–31656.

    Article  PubMed  CAS  Google Scholar 

  44. Linseman DA, Hofmann F, Fisher SK. A role for the small molecular weight GTPases, Rho and Cdc42, in muscarinic receptor signaling to focal adhesion kinase. J Neurochem. 2000;74:2010–2020.

    Article  PubMed  CAS  Google Scholar 

  45. Lee J, Jung ID, Chang WK, et al. p85 beta-PIX is required for cell motility through phosphorylations of focal adhesion kinase and p38 MAP kinase. Exp Cell Res. 2005;307:315–328.

    Article  PubMed  CAS  Google Scholar 

  46. 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;269:9345–9351.

    PubMed  CAS  Google Scholar 

  47. Salazar EP, Hunger-Glaser I, Rozengurt E. Dissociation of focal adhesion kinase and paxillin tyrosine phosphorylation induced by bombesin and lysophosphatidic acid from epidermal growth factor receptor transactivation in Swiss 3T3 cells. J Cell Physiol. 2003;194:314–324.

    Article  PubMed  CAS  Google Scholar 

  48. Leopoldt D, Yee HF Jr, Saab S, et al. Tyrosine phosphorylation of p125(Fak), p130(Cas), and paxillin does not require extracellular signal-regulated kinase activation in Swiss 3T3 cells stimulated by bombesin or platelet-derived growth factor. J Cell Physiol. 2000;183:208–220.

    Article  PubMed  CAS  Google Scholar 

  49. Chrzanowska-Wodnicka M, Burridge K. Tyrosine phosphorylation is involved in reorganization of the actin cytoskeleton in response to serum or LPA stimulation. J Cell Sci. 1994;107:3643–3654.

    PubMed  CAS  Google Scholar 

  50. Barry ST, Critchley DR. The RhoA-dependent assembly of focal adhesions in Swiss 3T3 cells is associated with increased tyrosine phosphorylation and the recruitment of both pp 125FAK and protein kinase C-delta to focal adhesions. J Cell Sci. 1994;107:2033–2045.

    PubMed  CAS  Google Scholar 

  51. Sawada K, Morishige K, Tahara M, et al. Alendronate inhibits lysophosphatidic acid-induced migration of human ovarian cancer cells by attenuating the activation of rho. Cancer Res. 2002;62:6015–6020.

    PubMed  CAS  Google Scholar 

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Acknowledgments

We sincerely thank Hong Xia (Key Laboratory of Hubei Province for Digestive System Disease, Wuhan, China) for his administrative support in this work. This work was supported by the National Natural Science Foundation of China (no. 81172350) and the Fundamental Research Funds for the Chinese Central Universities (no. 201130202020016 and no. 2012302020214).

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Authors and Affiliations

  1. Department of Gastroenterology, Renmin Hospital of Wuhan University, Number 238, Jiefang Road, Wuhan, 430060, Hubei Province, China

    Yan Liao, Ganggang Mu, Lingli Zhang, Wei Zhou, Jun Zhang & Honggang Yu

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Correspondence to Honggang Yu.

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Liao, Y., Mu, G., Zhang, L. et al. Lysophosphatidic Acid Stimulates Activation of Focal Adhesion Kinase and Paxillin and Promotes Cell Motility, via LPA1–3, in Human Pancreatic Cancer. Dig Dis Sci 58, 3524–3533 (2013). https://doi.org/10.1007/s10620-013-2878-4

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