NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
| Autor: | Silvio Holzner, Georg Krupitza, Katharina Viola, Nicole Huttary, Sigurd Krieger, Daniel Senfter, José Basílio, Serena Stadler, Daniela Milovanovic, Ingrid Simonitsch-Klupp, Chi Huu Nguyen, Walter Jäger, Rainer de Martin |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Pathology
medicine.medical_specialty NFKB1 RELA Breast Neoplasms Transfection chemistry.chemical_compound Paracrine signalling Cell Movement Cell Line Tumor Spheroids Cellular Paracrine Communication Basic Helix-Loop-Helix Transcription Factors Medicine Humans Receptor PAR-1 skin and connective tissue diseases business.industry Arabidopsis Proteins RELB NF-kappa B Endothelial Cells NF-κB Cell migration PAR1 Lymphatic system Oncology chemistry lymph endothelial cell migration Cancer cell Cancer research MCF-7 Cells Female Matrix Metalloproteinase 1 business MMP1 Intracellular Research Paper |
| Zdroj: | Oncotarget |
| ISSN: | 1949-2553 |
| Popis: | RELA, RELB, CREL, NFKB1 and NFKB2, and the upstream regulators NEMO and NIK were knocked-down in lymph endothelial cells (LECs) and in MDA-MB231 breast cancer spheroids to study the contribution of NF-κB in vascular barrier breaching. Suppression of RELA, NFKB1 and NEMO inhibited "circular chemo-repellent induced defects" (CCIDs), which form when cancer cells cross the lymphatic vasculature, by ~20-30%. Suppression of RELB, NFKB2 and NIK inhibited CCIDs by only ~10-15%. In MDA-MB231 cells RELA and NFKB1 constituted MMP1 expression, which caused the activation of PAR1 in adjacent LECs. The knock-down of MMP1 in MDA-MB231 spheroids and pharmacological inhibition of PAR1 in LECs inhibited CCID formation by ~30%. Intracellular Ca(2+) release in LECs, which was induced by recombinant MMP1, was suppressed by the PAR1 inhibitor SCH79797, thereby confirming a functional intercellular axis: RELA/NFKB1 - MMP1 (MDA-MB231) - PAR1 (LEC). Recombinant MMP1 induced PAR1-dependent phosphorylation of MLC2 and FAK in LECs, which is indicative for their activity and for directional cell migration such as observed during CCID formation. The combined knock-down of the NF-κB pathways in LECs and MDA-MB231 spheroids inhibited CCIDs significantly stronger than knock-down in either cell type alone. Also the knock-down of ICAM-1 in LECs (a NF-κB endpoint with relevance for CCID formation) and knock-down of MMP1 in MDA-MB231 augmented CCID inhibition. This evidences that in both cell types NF-κB significantly and independently contributes to tumour-mediated breaching of the lymphatic barrier. Hence, inflamed tumour tissue and/or vasculature pose an additional threat to cancer progression. |
| Databáze: | OpenAIRE |
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