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Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells

PURPOSE: Although neutrophil extracellular traps (NETs) are present in various tumors, their roles in tumor biology have not been clarified yet. In this study, we examined how NETs affect the pharmacokinetics and effects of doxorubicin (DOX). METHODS: NETs were generated by neutrophils stimulated wi...

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Autores principales: Tamura, Kohei, Miyato, Hideyo, Kanamaru, Rihito, Sadatomo, Ai, Takahashi, Kazuya, Ohzawa, Hideyuki, Koyanagi, Takahiro, Saga, Yasushi, Takei, Yuji, Fujiwara, Hiroyuki, Lefor, Alan Kawarai, Sata, Naohiro, Kitayama, Joji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9218137/
https://www.ncbi.nlm.nih.gov/pubmed/35756123
http://dx.doi.org/10.1016/j.heliyon.2022.e09730
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author Tamura, Kohei
Miyato, Hideyo
Kanamaru, Rihito
Sadatomo, Ai
Takahashi, Kazuya
Ohzawa, Hideyuki
Koyanagi, Takahiro
Saga, Yasushi
Takei, Yuji
Fujiwara, Hiroyuki
Lefor, Alan Kawarai
Sata, Naohiro
Kitayama, Joji
author_facet Tamura, Kohei
Miyato, Hideyo
Kanamaru, Rihito
Sadatomo, Ai
Takahashi, Kazuya
Ohzawa, Hideyuki
Koyanagi, Takahiro
Saga, Yasushi
Takei, Yuji
Fujiwara, Hiroyuki
Lefor, Alan Kawarai
Sata, Naohiro
Kitayama, Joji
author_sort Tamura, Kohei
collection PubMed
description PURPOSE: Although neutrophil extracellular traps (NETs) are present in various tumors, their roles in tumor biology have not been clarified yet. In this study, we examined how NETs affect the pharmacokinetics and effects of doxorubicin (DOX). METHODS: NETs were generated by neutrophils stimulated with phorbol 12-myristate 13-acetate (PMA) or lipopolysaccharide (LPS). DOX was added to NETs and their distribution was observed under fluorescein microscopy, and the diffusion of DOX through 3 μM pores from lower to upper chambers was evaluated with a fluorescence-based assay. Ovarian cancer cells, KOC-2S and SKOV3, were embedded in collagen gel droplets and cultured in 3D way and their apoptosis was examined with flow cytometry. RESULTS: DOX was mostly co-localized with NETs. The transfer of DOX to upper chambers increased over time, which was significantly decreased by the presence of neutrophils stimulated with PMA or LPS in the lower chamber. DOX outside of the gel increased the rates of annexin V (+) apoptotic cells, which were significantly reduced by the addition of LPS-stimulated neutrophils in media both in KOC-2S and SKOV3. The reduced diffusion and apoptosis were mostly restored by the destruction of the NETs structure with 1000 u/ml DNAse I. CONCLUSION: NETs efficiently trap and inhibit the diffusion of DOX which may attenuate its ability to induce apoptosis of ovarian cancer cells. Degradation of NETs with DNAse I may augment the response of ovarian cancer to DOX.
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spelling pubmed-92181372022-06-24 Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells Tamura, Kohei Miyato, Hideyo Kanamaru, Rihito Sadatomo, Ai Takahashi, Kazuya Ohzawa, Hideyuki Koyanagi, Takahiro Saga, Yasushi Takei, Yuji Fujiwara, Hiroyuki Lefor, Alan Kawarai Sata, Naohiro Kitayama, Joji Heliyon Research Article PURPOSE: Although neutrophil extracellular traps (NETs) are present in various tumors, their roles in tumor biology have not been clarified yet. In this study, we examined how NETs affect the pharmacokinetics and effects of doxorubicin (DOX). METHODS: NETs were generated by neutrophils stimulated with phorbol 12-myristate 13-acetate (PMA) or lipopolysaccharide (LPS). DOX was added to NETs and their distribution was observed under fluorescein microscopy, and the diffusion of DOX through 3 μM pores from lower to upper chambers was evaluated with a fluorescence-based assay. Ovarian cancer cells, KOC-2S and SKOV3, were embedded in collagen gel droplets and cultured in 3D way and their apoptosis was examined with flow cytometry. RESULTS: DOX was mostly co-localized with NETs. The transfer of DOX to upper chambers increased over time, which was significantly decreased by the presence of neutrophils stimulated with PMA or LPS in the lower chamber. DOX outside of the gel increased the rates of annexin V (+) apoptotic cells, which were significantly reduced by the addition of LPS-stimulated neutrophils in media both in KOC-2S and SKOV3. The reduced diffusion and apoptosis were mostly restored by the destruction of the NETs structure with 1000 u/ml DNAse I. CONCLUSION: NETs efficiently trap and inhibit the diffusion of DOX which may attenuate its ability to induce apoptosis of ovarian cancer cells. Degradation of NETs with DNAse I may augment the response of ovarian cancer to DOX. Elsevier 2022-06-15 /pmc/articles/PMC9218137/ /pubmed/35756123 http://dx.doi.org/10.1016/j.heliyon.2022.e09730 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Tamura, Kohei
Miyato, Hideyo
Kanamaru, Rihito
Sadatomo, Ai
Takahashi, Kazuya
Ohzawa, Hideyuki
Koyanagi, Takahiro
Saga, Yasushi
Takei, Yuji
Fujiwara, Hiroyuki
Lefor, Alan Kawarai
Sata, Naohiro
Kitayama, Joji
Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells
title Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells
title_full Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells
title_fullStr Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells
title_full_unstemmed Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells
title_short Neutrophil extracellular traps (NETs) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells
title_sort neutrophil extracellular traps (nets) reduce the diffusion of doxorubicin which may attenuate its ability to induce apoptosis of ovarian cancer cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9218137/
https://www.ncbi.nlm.nih.gov/pubmed/35756123
http://dx.doi.org/10.1016/j.heliyon.2022.e09730
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