Upregulation of the S1P3receptor in metastatic breast cancer cells increases migration and invasion by induction of PGE2and EP2/EP4activation.
Breast cancer is one of the most common and devastating malignancies among women worldwide. Recent evidence suggests that malignant progression is also driven by processes involving the sphingolipid molecule sphingosine 1-phosphate (S1P) and its binding to cognate receptor subtypes on the cell surface. To investigate the effect of this interaction on the metastatic phenotype, we used the breast cancer cell line MDA-MB-231 and the sublines 4175 and 1833 derived from lung and bone metastases in nude mice, respectively. In both metastatic cell lines expression of the S1P3receptor was strongly upregulated compared to the parental cells and correlated with higher S1P-induced intracellular calcium ([Ca2+]i), higher cyclooxygenase (COX)-2 and microsomal prostaglandin (PG) E2synthase expression, and consequently with increased PGE2synthesis. PGE2synthesis was decreased by antagonists and siRNA against S1P3and S1P2. Moreover, in parental MDA-MB-231 cells overexpression of S1P3by cDNA transfection also increased PGE2synthesis, but only after treatment with the DNA methyltransferase inhibitor 5-aza-2-deoxycytidine, indicating reversible silencing of the COX-2 promoter. Functionally, the metastatic sublines showed enhanced migration and Matrigel invasion in adapted Boyden chamber assays, which further increased by S1P stimulation. This response was abrogated by either S1P3antagonism, COX-2 inhibition or PGE2receptor 2 (EP2) and 4 (EP4) antagonism, but not by S1P2antagonism. Our data demonstrate that in breast cancer cells overexpression of S1P3and its activation by S1P has pro-inflammatory and pro-metastatic potential by inducing COX-2 expression and PGE2signaling via EP2and EP4.
Institute of Pharmacology, University of Bern, Inselspital, INO-F, CH-3011 Bern, Switzerland.
Iuliia
I
Filipenko
Pharmazentrum Frankfurt/ZAFES, Klinikum der Goethe Universität Frankfurt am Main, Theodor Stern Kai 7, D-60590 Frankfurt am Main, Germany.
Stephanie
S
Schwalm
Institute of Pharmacology, University of Bern, Inselspital, INO-F, CH-3011 Bern, Switzerland.
Luca
L
Reali
Pharmazentrum Frankfurt/ZAFES, Klinikum der Goethe Universität Frankfurt am Main, Theodor Stern Kai 7, D-60590 Frankfurt am Main, Germany.
Josef
J
Pfeilschifter
PIQUR Therapeutics AG, Hochbergstrasse 60C, CH-4057 Basel, Switzerland.
Doriano
D
Fabbro
Institute of Pharmacology, University of Bern, Inselspital, INO-F, CH-3011 Bern, Switzerland. Electronic address: huwiler@pki.unibe.ch.
Andrea
A
Huwiler
Institute of Pharmacology, University of Bern, Inselspital, INO-F, CH-3011 Bern, Switzerland. Electronic address: uwe.zangemeister@pki.unibe.ch.
Uwe
U
Zangemeister-Wittke
D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D002118:Calcium / Q000378:metabolism; D000068579:Celecoxib / Q000494:pharmacology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism* / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008246:Lysophospholipids / Q000494:pharmacology; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D000071876:Prostaglandin-E Synthases / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D049349:Receptors, Lysosphingolipid / Q000235:genetics / Q000378:metabolism*; D058307:Receptors, Prostaglandin E, EP2 Subtype / Q000235:genetics / Q000378:metabolism*; D058309:Receptors, Prostaglandin E, EP4 Subtype / Q000235:genetics / Q000378:metabolism*; D013110:Sphingosine / Q000031:analogs & derivatives / Q000494:pharmacology; D013997:Time Factors; D015854:Up-Regulation / Q000187:drug effects
D008246:Lysophospholipids; D012333:RNA, Messenger; D049349:Receptors, Lysosphingolipid; D058307:Receptors, Prostaglandin E, EP2 Subtype; D058309:Receptors, Prostaglandin E, EP4 Subtype; C060506:sphingosine 1-phosphate; D051546:Cyclooxygenase 2; C000605862:PTGES protein, human; D000071876:Prostaglandin-E Synthases; D000068579:Celecoxib; D015232:Dinoprostone; D013110:Sphingosine; D002118:Calcium
Breast cancer metastasis; Cyclooxygenase; Prostaglandin E(2); Prostaglandin E(2) receptor; Sphingosine 1-phosphate; Sphingosine 1-phosphate receptor 3