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Int J Mol Sci. 2018 Aug 9;19(8). pii: E2350. doi: 10.3390/ijms19082350.

Neutrophil Extracellular Traps as an Adhesion Substrate for Different Tumor Cells Expressing RGD-Binding Integrins.

Author information

1
Dipartimento di Medicina Clinica e Chirurgia, Università degli Studi di Napoli "Federico II", Via S. Pansini 5, 80131 Naples, Italy. marcello.monti@unina.it.
2
Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche, 80145 Naples, Italy. viviana.derosa@ibb.cnr.it.
3
Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche, 80145 Naples, Italy. francesca.iommelli@ibb.cnr.it.
4
Dipartimento di Oncologia Sperimentale, IRCCS Istituto Nazionale Tumori "Fondazione G. Pascale", 80145 Naples, Italy. m.carriero@istitutotumori.na.it.
5
Dipartimento di Scienze Biomediche Avanzate, Università degli Studi di Napoli "Federico II", Via S. Pansini 5, 80145 Naples, Italy. cr.terlizzi88@gmail.com.
6
Dipartimento di Oncologia Sperimentale, IRCCS Istituto Nazionale Tumori "Fondazione G. Pascale", 80145 Naples, Italy. r.camerlingo@istitutotumori.na.it.
7
Istituto di Genetica e Biofisica, Consiglio Nazionale delle Ricerche, 80131 Naples, Italy. stefania.belli@igb.cnr.it.
8
Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche, 80145 Naples, Italy. rosa.fonti@ibb.cnr.it.
9
Dipartimento di Medicina Clinica e Chirurgia, Università degli Studi di Napoli "Federico II", Via S. Pansini 5, 80131 Naples, Italy. diminno@unina.it.
10
Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche, 80145 Naples, Italy. delvecc@unina.it.
11
Dipartimento di Scienze Biomediche Avanzate, Università degli Studi di Napoli "Federico II", Via S. Pansini 5, 80145 Naples, Italy. delvecc@unina.it.

Abstract

Neutrophil extracellular traps (NETs), in addition to their function as a host defense mechanism, play a relevant role in thrombus formation and metastatic dissemination of cancer cells. Here we screened different cancer cell lines endogenously expressing a variety of integrins for their ability to bind to NETs. To this end, we used NETs isolated from neutrophil-like cells as a substrate for adhesion assays of HT1080, U-87 MG, H1975, DU 145, PC-3 and A-431 cells. Levels of α5, αIIb, αv, β1, β3 and β5 chains were determined by western blot analysis in all cell lines and levels of whole integrins on the plasma membrane were assessed by fluorescence-activated cell sorting (FACS) analysis. We found that high levels of α5β1, αvβ3 and αvβ5 enhance cell adhesion to NETs, whereas low expression of α5β1 prevents cell attachment to NETs. Excess of cyclic RGD peptide inhibited cell adhesion to NETs by competing with fibronectin within NETs. The maximal reduction of such adhesion was similar to that obtained by DNase 1 treatment causing DNA degradation. Our findings indicate that NETs from neutrophil-like cells may be used as a substrate for large screening of the adhesion properties of cancer cells expressing a variety of RGD-binding integrins.

KEYWORDS:

cancer metastasis; cell adhesion; fibronectin; integrins; neutrophil extracellular traps

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