S1P stimulates chemotactic migration and invasion in OVCAR3 ovarian cancer cells

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Abstract

OVCAR3 ovarian cancer cells express three sphingosine 1-phosphate (S1P) receptors, S1P1, S1P2, and S1P3, but not S1P4. Stimulation of OVCAR3 cells with S1P induced intracellular calcium increases, which were partly inhibited by VPC 23019 (an S1P1/3 antagonist). S1P-induced calcium increases were mediated by phospholipase C and pertussis toxin (PTX)-sensitive G-proteins in OVCAR3 cells. S1P stimulated extracellular signal-regulated kinase, p38 kinase, and Akt which were inhibited by PTX. S1P-stimulated chemotactic migration of OVCAR3 cells in a PTX-sensitive manner, indicating crucial role of Gi protein(s) in the process. S1P-induced chemotactic migration of OVCAR3 cells was completely inhibited by LY294002 and SB203580. Pretreatment of VPC 23019 (an S1P1/3 antagonist) completely inhibited S1P-induced chemotaxis. S1P also induced invasion of OVCAR3 cells, which was also inhibited by VPC 23019. Taken together, this study suggests that S1P stimulate chemotactic migration and cellular invasion, and VPC 23019-sensitive S1P receptor(s) might be involved in the processes.

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Materials and methods

Reagents and cell culture. S1P and VPC 23019 were purchased from Avanti Polar Lipids, Inc. (Alabaster, Alabama). The reverse transcription-polymerase chain reaction kit and fetal bovine serum (FBS) were purchased from Invitrogen Corporation (Carlsbad, CA). Phospho-extracellular signal-regulated protein kinase (ERK)1/2, phospho-p38 kinase, and ERK2 antibodies from New England Biolabs (Beverly, MA). LY294002, U-73122, U-73343, PD98059, and SB203580 were obtained from Calbiochem (San Diego, CA).

Functional expression of S1P receptors in OVCAR3 human ovarian cancer cells

We investigated whether cell surface receptors for S1P are expressed on OVCAR3 human ovarian cancer cells. To determine which S1P receptor isoforms are expressed on OVCAR3 human ovarian cancer cells, we analyzed the mRNA expressions of different S1P receptors by semi-quantitative RT-PCR. As shown in Fig. 1A, OVCAR3 human ovarian cancer cells expressed three isoforms of S1P receptor, namely, S1P1, S1P2, and S1P3. We were unable to detect S1P4 expression (Fig. 1A).

Activation of some

Discussion

Revealing extracellular factors and their target receptors which involve in the regulation of ovarian cancer cell migration, and invasion has been an important issue for the development of drugs for ovarian cancer therapy. Previously several groups have reported that various extracellular stimuli are involved in the regulation of ovarian cancer cell chemotactic migration and invasion [20], [23]. However, the role of S1P and its receptors in ovarian cancer cell chemotaxis and invasion has not

Acknowledgments

This work was supported by a Grant 02-PJ2-PG1-CH16-0002 from the Korea Health 21 R&D Project, Ministry of Health & Welfare, Republic of Korea, the Korea Science and Engineering Foundation through the Medical Science and Engineering Research Center for Cancer Molecular Therapy at Dong-A University, and the Korea Science and Engineering Foundation Grant (R01-2005-000-10011-02005).

References (27)

  • H.Y. Lee et al.

    A small compound that inhibits tumor necrosis factor-alpha-induced matrix metalloproteinase-9 upregulation

    Biochem. Biophys. Res. Commun.

    (2005)
  • S. Siehler et al.

    Pathways of transduction engaged by sphingosine 1-phosphate through G protein-coupled receptors

    Biochim. Biophys. Acta

    (2002)
  • D.Y. Noh et al.

    Phosphoinositide-specific phospholipase C and mitogenic signaling

    Biochim. Biophys. Acta

    (1995)

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