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Supplemental Figure 2 from Neutrophils Increase Oral Squamous Cell Carcinoma Invasion through an Invadopodia-Dependent Pathway

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posted on 2023-04-03, 23:01 authored by Judah E. Glogauer, Chun X. Sun, Grace Bradley, Marco A.O. Magalhaes

A- Live cell imaging of neutrophil and UMSCC47 co-cultures showing the intimate contact between cancer cells, matrix and neutrophils. The arrows point to neutrophils in close contact with cancer cells and matrix. B- Neutrophil culture showing the neutrophil-mediated matrix degradation. C- Neutrophil and UMSCC47 co-culture showing the dot-like invadopodia-mediated degradation and the neutrophil-mediated degradation. Neutrophils are washed away during fixation and therefore cannot be seen in these images. Cancer cells migrate during the 24h of the experiments and can be seen close to or on top of neutrophil degraded areas. Note that degradation "dots" caused by cancer cells are still visible under the neutrophil-mediated degradation.

History

ARTICLE ABSTRACT

Neutrophils have recently been shown to promote invasion and correlate with a poor prognosis in different cancers, including head and neck squamous cell carcinomas. In this study, we analyze the effects of neutrophils in the invasion of oral squamous cell carcinoma (OSCC) using a combination of conditioned media, direct and indirect coculture of human peripheral blood neutrophils, and UMSCC47 cells (OSCC cell line). Invasion and matrix degradation were determined using a modified in vitro invasion assay and an invadopodia assay, respectively. UMSCC47 and neutrophil cocultures or conditioned media from cocultures increased UMSCC47 invasion, invadopodia formation, and matrix degradation. Further analysis revealed an increase in TNFα and IL8 in supernatants of cocultures compared with neutrophil or UMSCC47 cultures alone and that inhibition of TNFα and IL8 significantly decreased OSCC invasion. Our results show that neutrophils increase the invasiveness of OSCC through the activation of invadopodia and matrix degradation, suggesting a paracrine activation loop between the two cells. Importantly, the presence of neutrophils in the oral environment may modulate the clinical behavior of OSCC. Cancer Immunol Res; 3(11); 1218–26. ©2015 AACR.