Up-regulation of Flotillin-2 Is Associated with Melanoma Progression and Modulates Expression of the Thrombin Receptor Protease Activated Receptor 1
Open Access
- 15 October 2004
- journal article
- Published by American Association for Cancer Research (AACR) in Cancer Research
- Vol. 64 (20), 7361-7369
- https://doi.org/10.1158/0008-5472.can-04-0823
Abstract
Flotillin 2 (flot-2) is a highly conserved protein isolated from caveolae/lipid raft domains that tether growth factor receptors linked to signal transduction pathways. Flot-2 protein and mRNA were increased in tumorigenic and metastatic melanoma cell lines in vitro, and the immunostaining intensity increased substantially across a tissue array of melanocytic lesions. Flot-2 transfection transformed SB2 melanoma cells from nontumorigenic, nonmetastatic to highly tumorigenic and metastatic in a nude mouse xenograft model. SB2 cells stably transfected with the flot-2 cDNA (SB2-flot)–2 cells proliferated faster in the absence of serum, and their migration through Matrigel was additionally enhanced by thrombin. When SB2-flot–2 cells were compared with SB2-vector–control cells on a cancer gene pathway array, SB2-flot–2 cells had increased expression of protease activated receptor 1 (PAR-1) mRNA, a transmembrane, G-protein–coupled receptor involved in melanoma progression. PAR-1 and flot-2 were coimmunoprecipitated from SB2-flot–2 cells. Up-regulation of PAR-1 was additionally confirmed in SB2-flot–2 cells and melanoma cell lines. SB2-flot–2 cells transfected with flot-2–specific small-interfering RNAs made substantially less flot-2 and PAR-1 mRNA. In conclusion, flot-2 overexpression is associated with melanoma progression, with increased PAR-1 expression, and with transformation of SB2 melanoma cells to a highly metastatic line. Flot-2 binds to PAR-1, a known upstream mediator of major signal transduction pathways implicated in cell growth and metastasis, and may thereby influence tumor progression.Keywords
This publication has 37 references indexed in Scilit:
- Membrane and raft association of reggie-1/flotillin-2: role of myristoylation, palmitoylation and oligomerization and induction of filopodia by overexpressionBiochemical Journal, 2004
- Adhesion-dependent Activation of the ERK1/2 Cascade Is By-passed in Melanoma CellsJournal of Biological Chemistry, 2003
- The Lipid Raft Microdomain-Associated Protein Reggie-1/ Flotillin-2 is Expressed in Human B Cells and Localized at the Plasma Membrane and Centrosome in PBMCsImmunobiology, 2002
- RhoA‐ and RhoD‐dependent regulatory switch of Gα subunit signaling by PAR‐1 receptors in cellular invasionThe FASEB Journal, 2002
- Suppression of matrix metalloproteinase-2 gene expression and invasion in human glioma cells by MMAC/PTENOncogene, 2001
- The thrombin receptor, PAR-1, causes transformation by activation of Rho-mediated signaling pathwaysOncogene, 2001
- Tumor Cell Invasion Is Promoted by Activation of Protease Activated Receptor-1 in Cooperation with the αvβ5 IntegrinPublished by Elsevier BV ,2001
- Flotillin and Epidermal Surface Antigen Define a New Family of Caveolae-associated Integral Membrane ProteinsPublished by Elsevier BV ,1997
- Epidermal Surface Antigen (M517S1) Is Highly Conserved between Mouse and HumanGenomics, 1995
- The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factorsCell, 1992