<<<<< << <<< er" content="American Association for Cancer Research" /> up/volpage/13/1019?iss=5" /> r-of-mouth models. GALR2 induced angiogenesis via p38-MAPK?mer-of-mouth models. GALR2 induced angiogenesis via p38-MAPK?mr-of-mouth models. GALR2 induced angiogenesis via p38-MAPK?mediated secretion of r-of-mouth models. GALR2 induced angiogenesis via p38-MAPK?mediated secretion of proangiogenic cytokines, VEGF, and interleur-of-mouth models. GALR2 induced angiogenesis via p38-MAPK?mediated secretion of proangiogenic cytokines, VEGF, and interleukin-6 (IL-6). Moreover, GALR2 activated small-GTP-protein, RAP1B, thereby inducing p38-mediated inactivation of tristetraprolin (TTP), which functions to destabilize cytokine transcripts. This resulted in enhanced secretion of proangiogenic cytokines and angiogenesis in vitro and in vivo . In SCCHN cells overexpressing GALR2, inactivation of TTP increased secretion of IL-6 and VEGF, whereas inhibition of p38 activated TTP and decreased cytokine secretion. Here, we report that GALR2 stimulates tumor angiogenesis in SCCHN via p38-mediatused to understand the key regulators. The <i>in vivo</i> impact of GALR2 on angiogenesis was investigated in mouse xenograft, chick chorioallantoic membrane, and the clinically relevant mouse orthotopic floor-of-mouth models. GALR2 induced angiogenesis via p38-MAPK?mediated secretion of proangiogenic cytokines, VEGF, and interleukin-6 (IL-6). Moreover, GALR2 activated small-GTP-protein, RAP1B, thereby inducing p38-mediated inactivation of tristetraprolin (TTP), which functions to destabilize cytokine transcripts. This resulted in enhanced secretion of proangiogenic cytokines and angiogenesis <i>in vitro</i> and <i>in vivo</i>. In SCCHN cells overexpressing GALR2, inactivation of TTP increased secretion of IL-6 and VEGF, whereas inhibition of p38 activated TTP and decreased cytokine secretion. Here, we report that GALR2 stimulates tumor angiogenesis in SCCHN via p38-mediated inhibition of TTP with resultant enhanced cytokine secretion. Given that p38 inhibitors are in clinical use for inflammatory disorders, GALR2/p38-mediated cytokine secretion may be an excellent target for new adjuvant therapy in SCCHN. <i>Mol Cancer Ther; 13(5); 1323?33. 2014 AACR</i>.</p>" /> used to understand the key regulators. The <i>in vivo</i> impact ofused to understand the key regulators. The <i>in vivo</i> impact of GALR2 used to understand the key regulators. The <i>in vivo<used to understand the key regulators. The <i>in vivo&lused to understand the key regulators. The <iused to understand the key regulators. The <i>in vivo</i> impact of GALR2 on angused to understand the key regulators. The <i>in vivo</i> impact of GALR2 on angiogenesis was inused to understand the key regulators. The <i>in vivo</i> impact of GALR2 on angiogenesiused to understand the key regulators. The <i>in vivo</i> impact of GALR2 on angiogeneused to understand the key regulators. The <i&gused to understand the ke" content="1538-8514" /> " content="1538-8514" /> iversity of Michigan Medical School, Ann Arbor, Michiganiversity of Michigan Medical School, Ann Arbor, Michigan; Department of Craniofacial Biology, The Medical University of South Carolina, Columbia, South Carolina" /> stry; Department of Pathology, University of Michigan Medical stry; Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan; Department of Craniofacial Biology, The Medical University of South Carolina, Columbia, South Carolina" /> tution" content="Authors' Affiliations: Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry; Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan; Department of Craniofacial Biology, The Medical University of South Carolina, Columbia, South Carolina" /> ent of Head and Neck Cancer and Its Inhibition By Chemopreventive Agents;citation_pages=153-164;citation_volume=10;citation_year=1999;citation_issue=2;citation_issn=1045-4411;citation_pmid=10759419;citation_doi=10.1177/10454411990100020301" /> urakis;citation_author=A. Sotiropoulou-Lontou;citation_title=Targeted therapy with bevacizumab (Avastin) for metastatic colorectal cancer.;citation_pages=710-714;citation_volume=13;citation_year=2011;citation_issue=10;citation_pmid=21975331;citation_doi=10.1007/s12094-011-0720-z" /> _author=JJ. Shacka;citation_title=Tristetraprolin Down-regulates Interleukin-8 and Vascular Endothelial Growth Factor in Malignant Glioma Cells;citation_pages=674-682;citation_volume=68;citation_year=2008;citation_issue=3;citation_issn=0008-5472;citation_pmid=18245466;citation_doi=10.1158/0008-5472.CAN-07-2751" /> has anti-proliferative effects in oral squamous cell carcinoma;citation_volume=280;citation_year=2005;citation_pmid=15767248;citation_doi=10.1074/jbc.M414589200" /> ll carcinoma.;citation_pages=132-142;citation_volume=48;citation_year=2009;citation_issue=2;citation_pmid=18973137;citation_doi=10.1002/gcc.20626" /> al in Low N-Stage Squamous Cell Carcinoma;citation_pages=3959-3969;n Cancer Res;citation_author=EA. Van Tubergen;citation_author=R. Banerjee;citation_author=M. Liu;citation_author=R. Vander Broek;citation_author=E. Light;citation_author=S. Kuo;citation_title=Inactivation or Loss of TTP Promotes Invasion in Head and Neck Cancer via Transcript Stabilization and Secretion of MMP9, MMP2, and IL-6;citation_volume=19;citation_year=2013;citation_pmid=23349315;citation_doi=10.1158/1078-0432.CCR-12-2927" /> n Cancer Res;citation_author=EA. Van Tubergen;citation_author=R. Banerjee;citation_author=M. Liu;citation_author=R. Vander Broek;citation_author=E. Light;citation_author=S. Kuo;citation_title=Inactivation or Loss of TTP Promotes Invasion in Head and Neck Cancer via Transcript Stabilization and Secretion of MMP9, MMP2, and IL-6;citation_volume=19;citation_year=2013;citation_pmid=23349315;citation_doi=10.1158/1078-0432.CCR-12-2927" /> e=Rap1GAP inhibits tumor growth in oropharyngeal squamous cell carcinoma.;citation_pages=585-596;citation_volume=168;citation_year=2006;citation_issue=2;citation_issn=0002-9440;citation_pmid=16436672;citation_doi=10.2353/ajpath.2006.050132" /> 8;citation_year=2006;citation_issue=3;citation_pmid=16611409;citation_doi=10.1593/neo.05691" /> and lethality in embryos lacking a single VEGF allele.;citation_pages=435-439;citation_volume=380;citation_year=1996;citation_issue=6573;citation_issn=0028-0836;citation_pmid=8602241;citation_doi=10.1038/380435a0" /> content="citation_journal_title=Oncogene;citation_journal_abbrev=Oncogene;citation_author=N. Wittau;citation_author=R. Grosse;citation_author=F. Kalkbrenner;citation_author=A. 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Amornphimoltham;citation_title=Decreased lymphangiogenesis and lymph node metastasis by mTOR inhibition in head and neck cancer;citation_volume=71;citation_year=2011;citation_pmid=21975930;citation_doi=10.1158/0008-5472.CAN-10-3192" /> s proliferation in squamous carcinoma cells: activation of the extracellular signal regulated kinase pathway and induction of cyclin-dependent kinase inhibitors.;citation_pages=5762-5771;citation_volume=26;citation_year=2007;citation_issue=39;citation_pmid=17384686;citation_doi=10.1038/sj.onc.1210384" /> n_pages=338-46;citation_volume=6;citation_year=2013;citation_pmid=23730414;citation_doi=10.1593/tlo.13115" /> n_pages=338-46;<link type="text/css" rel="stylesheet" href="//mct.aacrjournals.org/sites/default/files/advagg_css/css__k4rrujIpcQouOYNFR--LXVAGmksNyOzxFeQGguag-Jc__i7Qnw6YFvAAEfiQwMErS76BYBKCHQGmP5SEwBspTeB8__rv6vj8L029q-VeAfCopTipIESoW6SpXaZvbMqVdrQG8.css" media="all"/>n<link type="text/css" rel="stylesheet" href="//mct.aacrjournals.org/sites/all/modules/highwire/highwire/highwire.style.highwire.css?p0k9mh" media="all"/>n<link type="text/css" rel="stylesheet" href="//mct.aacrjournals.org/sites/default/files/advagg_css/css__KPgiwEdjo5O9HdRryHqQs1TpsV-PeBb9B7sGBvwM9FQ__opzZWgPNgQzoEgfATRS5qCKwBw_6XcNjDHoDVOzQv8M__rv6vj8L029q-VeAfCopTipIESoW6SpXaZvbMqVdrQG8.css" media="all"/>n<link type="text/css" rel="stylesheet" href="//cdn.jsdelivr.net/qtip2/2.2.1/jquery.qtip.min.css" media="all"/>n<link type="text/css" rel="stylesheet" href="//mct.aacrjournals.org/sites/default/files/advagg_css/css__DDPP3F46sogJvfA4OaTajx6FS4_4WSmX7LaDAqF4PMg__DQjj5IAP37g6XxBxp3Ws2AFF73y9tFAb8edIkA0-Gnw__rv6vj8L029q-VeAfCopTipIESoW6SpXaZvbMqVdrQG8.css" media="all"/>n_n_pages=338-46;citation_volume=6;citation_year=2013;citation_pmid=23730414;citation_doi=10.1593/tlo.13115" /> <title>The G Protein?Coupled Receptor GALR2 Promotes Angiogenesis in Head and Neck Cancer | Molecular Cancer Therapeutics nAfAfCopTipIESoW6SpXaZvbMqVdrQG8.css" media="all" /> 8L029q-VeAfCopTipIESoW6S88L029q-VeAfCopTip8L029hhhhhh> <> >> <> > <> >> <> >> <> aaaaaa ee-advere-advertise-advertie-advere-adverte-advere-adverte-advere-advee-advertising-top clearfix mobile-hidden container-30">
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Conditioned medium

Conditioned medium (CM) was collected as described (6). Briefly, SCCHN cells at approximately 60% to 70% confluence were cul <

Conditioned medium

Conditio

Conditioned medium

Conditioned medium (CM) was collected as described (Conditioned mediumC

Conditioned medium

Condit

Conditioned medium

Conditioned medium (CM) was collected as described (6). Briefly, SCCHN cells at approximately 60% to 70% confluence were cultured in Dulbecco's Modified Eagle Medium (DMEM) without supplements. After 24 hours, the medium was centrifuged and the supernorseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch) were visualized by SuperSignal Substrate (Pierce).orseorseradorseradish peroxidase-conjugated secorseradish peroxidase-corseraorseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch) were visualized by SuperSignal Substrate (Pierce).

Immunoprecipitation

ororseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch) were visualized by SuperSignal Substrate (Pierce).

Immunoprecipitation

TTP was immunoprecipitated with TTP antibody (Santa Cruz Biotechnology) crosslinked to Amino-Link Plus Coupling Resin (Pierce) supplemented with protease (Roche) and phosphatase (Sigma) inhibitors.

ELISA

ororseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch) were visualized by SuperSignal Substrate (Pierce).

In vivo studies and immunohistochemistr hours hours and incubated with 10 ?mol/L of hour hours and incubated with 10 ?mol/L of SB203580 (Promega) for 1 hour.

In vivo h hours and incubated with 10 ?mol/L of SB203580 (Promega) for 1 hour.

In vivo studies and immunohistochemistry

All in vivo studies were done according to Univers hours hours and incubated with 10 ?mol/L of SB203580 (Promega) for 1 hour.

h hours and incubated with 10 ?mol/L of SB203580 (Promega) for 1 hour.

h hours and incubated with 10 hou hours hours and incubated with 10 ?mol/L o hours and incu hours hours and incubated with 10 ?mol/L of SB203580 (Promega) for 1 hour.

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Previously, we showed that overexpression of GALR2 in multiple SCCHN cell lines induces proliferation and sis

Previously, we showedisis

Previously, we showed that overexpression of GALR2 in multiple SCCHN cell lines induces proliferation and survival in vitro<is

is

Previously, we showed that overexpression of GALR2 in multiple SCCHN cell lines induces proliferation and survival in vitro and aggressive tumors in the mouse (16). In the present study, we show that tumors generated from UM-SCC-1-GALR2 cells are more vasculaisis

Previously, we showed that overexpression of GALR2 in multiple SCCHN cell lines induces proliferation and survival in vitro and aggressive tumors in the mouse (

Previously, we showis

Previousis

Previously, we is

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