Localization of uPAR and MMP-9 in lipid rafts is critical for migration, invasion and angiogenesis in human breast cancer cells
Open Access
- 24 November 2010
- journal article
- Published by Springer Science and Business Media LLC in BMC Cancer
- Vol. 10 (1), 647
- https://doi.org/10.1186/1471-2407-10-647
Abstract
Background: uPAR and MMP-9, which play critical roles in tumor cell invasion, migration and angiogenesis, have been shown to be associated with lipid rafts. Methods: To investigate whether cholesterol could regulate uPAR and MMP-9 in breast carcinoma, we used MβCD (methyl beta cyclodextrin, which extracts cholesterol from lipid rafts) to disrupt lipid rafts and studied its effect on breast cancer cell migration, invasion, angiogenesis and signaling. Results: Morphological evidence showed the association of uPAR with lipid rafts in breast carcinoma cells. MβCD treatment significantly reduced the colocalization of uPAR and MMP-9 with lipid raft markers and also significantly reduced uPAR and MMP-9 at both the protein and mRNA levels. Spheroid migration and invasion assays showed inhibition of breast carcinoma cell migration and invasion after MβCD treatment. In vitro angiogenesis studies showed a significant decrease in the angiogenic potential of cells pretreated with MβCD. MβCD treatment significantly reduced the levels of MMP-9 and uPAR in raft fractions of MDA-MB-231 and ZR 751 cells. Phosphorylated forms of Src, FAK, Cav, Akt and ERK were significantly inhibited upon MβCD treatment. Increased levels of soluble uPAR were observed upon MβCD treatment. Cholesterol supplementation restored uPAR expression to basal levels in breast carcinoma cell lines. Increased colocalization of uPAR with the lysosomal marker LAMP1 was observed in MβCD-treated cells when compared with untreated cells. Conclusion: Taken together, our results suggest that cholesterol levels in lipid rafts are critical for the migration, invasion, and angiogenesis of breast carcinoma cells and could be a critical regulatory factor in these cancer cell processes mediated by uPAR and MMP-9.Keywords
This publication has 52 references indexed in Scilit:
- Src Family Kinases Accelerate Prolactin Receptor Internalization, Modulating Trafficking and Signaling in Breast Cancer CellsMolecular Endocrinology, 2009
- Clathrin and LRP-1-Independent Constitutive Endocytosis and Recycling of uPARPLOS ONE, 2008
- Lipid Raft Disruption by Cholesterol Depletion Enhances Influenza A Virus Budding from MDCK CellsJournal of Virology, 2007
- Lipid Rafts of Primary Endothelial Cells Are Essential for Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8-Induced Phosphatidylinositol 3-Kinase and RhoA-GTPases Critical for Microtubule Dynamics and Nuclear Delivery of Viral DNA but Dispensable for Binding and EntryJournal of Virology, 2007
- Retracted: RNAi‐mediated downregulation of urokinase plasminogen activator receptor and matrix metalloprotease‐9 in human breast cancer cells results in decreased tumor invasion, angiogenesis and growthInternational Journal of Cancer, 2007
- Cholesterol inhibits MMP‐9 expression in human epidermal keratinocytes and HaCaT cellsFEBS Letters, 2007
- RNA interference–mediated simultaneous down-regulation of urokinase-type plasminogen activator receptor and cathepsin B induces caspase-8–mediated apoptosis in SNB19 human glioma cellsMolecular Cancer Therapeutics, 2006
- Unbiased quantitative proteomics of lipid rafts reveals high specificity for signaling factorsProceedings of the National Academy of Sciences, 2003
- Role of sphingomyelinase and ceramide in modulating rafts: do biophysical properties determine biologic outcome?FEBS Letters, 2002
- Modulation of ?5?1 integrin functions by the phospholipid and cholesterol contents of cell membranesJournal of Cellular Biochemistry, 2000