Crosstalk between protease-activated receptor 1 and platelet-activating factor receptor regulates melanoma cell adhesion molecule (MCAM/MUC18) expression and melanoma metastasis.
The cellular and molecular pathways that regulate platelet activation, blood coagulation, and inflammation are emerging as critical players in cancer progression and metastasis. Here, we demonstrate a novel signaling mechanism whereby protease-activated receptor 1 (PAR1) mediates expression of melanoma cell adhesion molecule MCAM/MUC18 (MUC18), a critical marker of melanoma metastasis, via activation of platelet-activating factor receptor (PAFR) and cAMP-responsive element-binding protein (CREB). We found that PAR1 silencing with small hairpin RNA inhibits MUC18 expression in metastatic melanoma cells by inhibiting CREB phosphorylation, activity, and binding to the MUC18 promoter. We further demonstrate that the PAF/PAFR pathway mediates MUC18 expression downstream of PAR1. Indeed, PAR1 silencing down-regulates PAFR expression and PAF production, PAFR silencing blocks MUC18 expression, and re-expression of PAFR in PAR1-silenced cells rescues MUC18 expression. We further demonstrate that the PAR1-PAFR-MUC18 pathway mediates melanoma cell adhesion to microvascular endothelial cells, transendothelial migration, and metastatic retention in the lungs. Rescuing PAFR expression in PAR1-silenced cells fully restores metastatic phenotype of melanoma, indicating that PAFR plays critical role in the molecular mechanism of PAR1 action. Our results link the two pro-inflammatory G-protein-coupled receptors, PAR1 and PAFR, with the metastatic dissemination of melanoma and suggest that PAR1, PAFR, and MUC18 are attractive therapeutic targets for preventing melanoma metastasis.
Melnikova
Vladislava O
VO
Department of Cancer Biology, The University of Texas M D Anderson Cancer Center, Houston, Texas 77030, USA.
Balasubramanian
Krishnakumar
K
Villares
Gabriel J
GJ
Dobroff
Andrey S
AS
Zigler
Maya
M
Wang
Hua
H
Petersson
Frederik
F
Price
Janet E
JE
Schroit
Alan
A
Prieto
Victor G
VG
Hung
Mien-Chie
MC
Bar-Eli
Menashe
M
D051929:CD146 Antigen / Q000378:metabolism; D050882:CREB-Binding Protein / Q000378:metabolism; D045744:Cell Line, Tumor; D020868:Gene Silencing; D006801:Humans; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology; D009362:Neoplasm Metastasis; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D044463:Receptor, PAR-1 / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D012878:Skin Neoplasms / Q000378:metabolism* / Q000473:pathology; D014157:Transcription Factors / Q000378:metabolism
D051929:CD146 Antigen; C497902:MCAM protein, human; D010980:Platelet Membrane Glycoproteins; D034741:RNA, Small Interfering; D044463:Receptor, PAR-1; D043562:Receptors, G-Protein-Coupled; D014157:Transcription Factors; C039351:platelet activating factor receptor; D050882:CREB-Binding Protein
Oncology. 2005;69(3):185-201
16138000
Cancer Metastasis Rev. 1999;18(3):345-57
10721489
Can J Biochem Physiol. 1959 Aug;37(8):911-7
13671378
Am J Pathol. 2008 Dec;173(6):1839-52
18988806
Curr Pharm Des. 2001 Nov;7(16):1615-26
11562302
J Exp Med. 2002 Jan 21;195(2):171-9
11805144
Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8205-9
7667269
Proc Natl Acad Sci U S A. 1968 Sep;61(1):46-52
5246932
J Biol Chem. 2003 Nov 21;278(47):46632-42
12975361
J Biol Chem. 1995 Jun 23;270(25):14928-34
7797472
Thromb Res. 1992 Nov 1;68(3):233-45
1281930
J Virol. 2003 Aug;77(16):8957-61
12885912
J Biol Chem. 2004 Jan 30;279(5):3497-508
14617636
Oncogene. 1997 Oct 23;15(17):2069-75
9366524
Cancer Res. 1997 Jun 1;57(11):2295-303
9187135
Nature. 2008 Apr 3;452(7187):654-8
18305483
Cancer Cell. 2006 Nov;10(5):355-62
17097558
Cell Tissue Res. 2009 Jan;335(1):249-59
18815812
Am J Pathol. 2000 Nov;157(5):1713-25
11073830
Cancer Res. 1995 Apr 15;55(8):1629-32
7712465
Nat Med. 1998 Aug;4(8):909-14
9701242
Nat Rev Mol Cell Biol. 2001 Aug;2(8):599-609
11483993
Blood. 1998 Nov 15;92(10):3694-700
9808563
Int J Cancer. 1991 Jan 21;47(2):227-37
1671030
Am J Pathol. 2007 May;170(5):1781-92
17456781
J Biol Chem. 2001 Apr 6;276(14):10952-62
11278329
Clin Cancer Res. 2003 Sep 15;9(11):4214-20
14519648
J Biol Chem. 1998 Oct 9;273(41):26852-6
9756930
N Engl J Med. 2008 Dec 18;359(25):2641-50
19092150
Lab Invest. 1991 Nov;65(5):525-31
1753702
Ann N Y Acad Sci. 2007 Jan;1096:86-8
17405919
Cancer Res. 1994 May 1;54(9):2514-20
8162602
Cancer Res. 2009 Aug 15;69(16):6730-7
19679555
Cancer Res. 1996 May 1;56(9):2218-23
8616875
Cancer Metastasis Rev. 2008 Mar;27(1):11-8
18066650
Int J Cancer. 1993 Apr 1;53(6):978-82
8473056
Cancer Res. 2004 Sep 15;64(18):6482-8
15374958
Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11023-7
10500117
Blood. 2008 Jan 1;111(1):190-9
17901245
Oncogene. 2001 Apr 12;20(16):1953-63
11360179
J Biol Chem. 2000 Nov 3;275(44):34672-80
10938273
Cancer Res. 2008 May 15;68(10):3978-84
18483284
J Biol Chem. 2009 Sep 18;284(38):26194-206
19632997
J Invest Dermatol. 2007 Feb;127(2):387-93
16946713
J Invest Dermatol. 2000 Aug;115(2):267-72
10951245
Urology. 2002 Nov;60(5):760-5
12429291
Nature. 2008 Jul 24;454(7203):436-44
18650914
Biotechniques. 2002 Oct;33(4):842-4, 846, 848-50
12398193
Br J Cancer. 1998 Jun;77(12):2069-75
9649116
Cancer Res. 2002 Sep 1;62(17):5106-14
12208768
J Biol Chem. 2006 Feb 3;281(5):2911-22
16306050
Cell. 2005 Feb 11;120(3):303-13
15707890
J Immunol. 1998 Aug 15;161(4):1954-61
9712066
J Exp Med. 2008 Aug 4;205(8):1739-46
18606855
J Pathol. 1999 Sep;189(1):4-11
10451481
Cancer Res. 2008 Nov 1;68(21):9078-86
18974154
Oncogene. 2001 Aug 2;20(34):4676-84
11498790
Semin Thromb Hemost. 2006 Apr;32 Suppl 1:61-8
16673267
Cancer Res. 1997 Sep 1;57(17):3835-40
9288796
Neoplasia. 2005 May;7(5):522-7
15967104
Cell. 2008 Apr 4;133(1):66-77
18394990