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. 2020 Jan-Dec;27(1):1073274820960473.
doi: 10.1177/1073274820960473.

Cancers Cells in Traps? The Pathways of NETs Formation in Response to OSCC in Humans-A Pilot Study

Marzena Garley  1 , Ewa Jabłońska  1 , Wojciech Miltyk  2 , Kamil Grubczak  3 , Arkadiusz Surażyński  4 , Wioletta Ratajczak-Wrona  1 , Małgorzata Grudzińska  5 , Kinga H Nowacka  1 , Marcin Moniuszko  3 , Jerzy A Pałka  4 , Jan Borys  6 , Dorota Dziemiańczyk-Pakieła  7
Affiliations

Cancers Cells in Traps? The Pathways of NETs Formation in Response to OSCC in Humans-A Pilot Study

Marzena Garley et al. Cancer Control. 2020 Jan-Dec.

Abstract

The aim of the experiment was to evaluate the process of neutrophil extracellular traps (NETs) formation in patients with oral squamous cell carcinoma (OSCC) in response to direct or indirect contact with SCC cells in comparison to results obtained in the cells of healthy subjects. To fulfill study objectives CAL 27 cell line and blood were obtained from cancer patients and control subjects. Parameters related to NETs formation were analyzed utilizing flow cytometry, fluorescence microscopy, and ELISA-type tests. The expression of selected phosphorylated proteins of the PI3K/Akt/PBK pathway in neutrophils was evaluated using the Western blot method. An increase in NETs formation was observed in a coculture of neutrophils with SCC cells, with the largest amount of NETs formed after stimulation with a supernatant obtained from the SCC culture. The enhanced process of NETs formation was accompanied by changes in the expression of proteins from the PI3K/Akt/PBK pathway. The obtained results prove the existence of interactions between neutrophils and cancer cells resulting in NETosis with the participation of the PI3K/Akt/PBK pathway in patients with OSCC.

Keywords: NETs; PI3K/Akt/PBK; SCC; neutrophils; oral cancer.

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Conflict of interest statement

Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
NETs under the microscope. Example of NETs image of coculture of neutrophils from a patient with cancer in response to SCC cells: in blue—DNA (Hoechst 33342), in red—MPO monoclonal antibodies labeled with fluorescein (Clone 8E6).
Figure 2.
Figure 2.
Representative results for MPO-positive neutrophils determined by flow cytometry.
Figure 3.
Figure 3.
Western blot results. The figure shows the results obtained by the Western blot method: A—sample scans of membrane samples from healthy people and patients with OSCC; B—column charts of average expression values of tested proteins together with SD and statistically significant differences: *—with control; a—between PMNs and PMNs+SCC cells; b—between PMNs and PMNs+Supernatant; c—between PMNs+SCC cells and PMNs+Supernatant.
See this image and copyright information in PMC

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