S1P regulation of ovarian carcinoma invasiveness

Smicun, Yoel; Reierstad, Scott; Wang, Feng-Qiang; Lee, Cathy; Fishman, David A.

doi:10.1016/j.ygyno.2006.06.036

Cited by 31 publications

(22 citation statements)

References 28 publications

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“…This may explain the different response of OVCA 429 and Dov13 cells to the PTX inhibition of the low-dose-S1P induced increase of gelatinase activity. We also reported that treatment with S1P alters the surface expression of S1P1, 2, and 3 in a concentration-dependent fashion [12]. As discussed, multiple S1P-receptor-coupled pathways converge at Rac, some that are Gi dependent and some that act through G12/13 or Gq [14].…”

Section: Discussionmentioning

confidence: 74%

“…We have previously demonstrated that the five known S1P specific receptors are differently expressed in the EOC cell lines DOV 13 and OVCA 429 [12]. This may explain the different response of OVCA 429 and Dov13 cells to the PTX inhibition of the low-dose-S1P induced increase of gelatinase activity.…”

Section: Discussionmentioning

confidence: 99%

“…For whole cell lysates, cells were washed with ice-cold PBS and then extracted with lysis buffer (Radioimmunoprecipitation assay, RIPA, buffer containing 2 mM PMSF, 2 mM Na 3 VO 4 , 2 mM NaF, and aprotinin, leupeptin and pepstatin of 2 µg/ml each). Cell surface proteins of treated cells were biotin labeled and isolated as described [12] using the Pierce Biotinylation kit (Rockford, IL). Briefly, cells in plates were labeled with Sulfo-NHS-SS-Biotin, quenched, scraped off plates, sonicated in lysis buffer, and affinity purified by Immobilized NeutrAvidin.…”

Section: Western Analysismentioning

confidence: 99%

“…While LPA has been extensively studied, less is known about the mechanisms and behaviors of S1P. Recently, we reported that S1P induces EOC invasiveness similar to LPA at low concentrations yet inhibits invasion at high concentrations [12]. The pathways leading to the concentration dependant effect are unknown.…”

Section: Introductionmentioning

confidence: 97%

“…There are five known SP1 receptors encoded by members of the endothelial differentiation gene (Edg) family, S1P [1][2][3][4][5] [9]. We have recently shown that S1P regulates transcription and surface presentation of its own receptors [12]. The G proteins that mediate the biologic effects of S1P are associated with particular Edg's, and pathways have been identified to all known S1P receptors through coupled G-Proteins, including the Gi subgroup [13].…”

Section: Introductionmentioning

confidence: 99%

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S1P induced changes in epithelial ovarian cancer proteolysis, invasion, and attachment are mediated by Gi and Rac

Devine

Smicun

Hope

et al. 2008

Gynecologic Oncology

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OBJECTIVES-We previously demonstrated that sphingosine 1-phosphate (S1P) bimodally regulates epithelial ovarian cancer (EOC) cell invasiveness: low-concentration S1P stimulates invasion similar to lysophophatidic acid (LPA), while high-concentration S1P inhibits invasion. In this study, we investigated the mechanisms through which S1P affects EOC cell proteolysis, invasion, and adhesion in two cultured epithelial ovarian cancer cell lines. METHODS-G-proteinGi was inhibited by pertussis toxin (PTX) and GTP binding protein Rac by NSC23766. S1P conditioned media of DOV13 and OVCA429 cells were evaluated via gel zymography, fluorometric gelatinase assay, urokinase plasminogen activator (uPA) activity assay, and Western Blot for MT1-MMP. Cell invasion was analyzed in Matrigel chambers. Membrane-N-cadherin was localized via fluorescence microscopy.RESULTS-Zymography revealed pro-MMP2 in conditioned media of EOC cells regardless of treatment. Gelatinase activity was increased by low-concentration S1P. In DOV13 cells this effect was Gi and Rac dependent. In all OVCA429 and control DOV13 cells, PTX enhanced gelatinolysis, suggesting an MMP2-inhibitory pathway via Gi. MT1-MMP was decreased Gidependently by high-concentration S1P. Rac inhibition significantly counteracted low-S1P enhancement and high-S1P reduction of DOV13 invasiveness; and uPA activity in conditioned media of invading cells correlated significantly. Immunohistochemistry revealed Gi-dependent clustering of membrane-N-cadherin in DOV13 cells treated with 0.5µM S1P or 10µM LPA.CONCLUSIONS-S1P influences EOC invasion by regulating ECM-proteolysis and cell-cell attachment via MMP2, uPA, and membrane-N-cadherin. Furthermore, this study illustrates that the net effect of S1P on each of these processes reflects a complex interplay of multiple GPCR pathways involving Gi and downstream Rac.

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Section: Discussionmentioning

confidence: 74%

Section: Discussionmentioning

confidence: 99%

Section: Western Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%