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Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines

Autophagy, a self-digestive system for cytoplasmic components, is required to maintain the amino acid pool for cellular homeostasis. We previously reported that the macrolide antibiotics azithromycin (AZM) and clarithromycin (CAM) have an inhibitory effect on autophagy flux, and they potently enhanc...

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Autores principales: Hirasawa, Kazuhiro, Moriya, Shota, Miyahara, Kana, Kazama, Hiromi, Hirota, Ayako, Takemura, Jun, Abe, Akihisa, Inazu, Masato, Hiramoto, Masaki, Tsukahara, Kiyoaki, Miyazawa, Keisuke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5158196/
https://www.ncbi.nlm.nih.gov/pubmed/27977675
http://dx.doi.org/10.1371/journal.pone.0164529
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author Hirasawa, Kazuhiro
Moriya, Shota
Miyahara, Kana
Kazama, Hiromi
Hirota, Ayako
Takemura, Jun
Abe, Akihisa
Inazu, Masato
Hiramoto, Masaki
Tsukahara, Kiyoaki
Miyazawa, Keisuke
author_facet Hirasawa, Kazuhiro
Moriya, Shota
Miyahara, Kana
Kazama, Hiromi
Hirota, Ayako
Takemura, Jun
Abe, Akihisa
Inazu, Masato
Hiramoto, Masaki
Tsukahara, Kiyoaki
Miyazawa, Keisuke
author_sort Hirasawa, Kazuhiro
collection PubMed
description Autophagy, a self-digestive system for cytoplasmic components, is required to maintain the amino acid pool for cellular homeostasis. We previously reported that the macrolide antibiotics azithromycin (AZM) and clarithromycin (CAM) have an inhibitory effect on autophagy flux, and they potently enhance the cytocidal effect of various anticancer reagents in vitro. This suggests that macrolide antibiotics can be used as an adjuvant for cancer chemotherapy. Since cancer cells require a larger metabolic demand than normal cells because of their exuberant growth, upregulated autophagy in tumor cells has now become the target for cancer therapy. In the present study, we examined whether macrolides exhibit cytotoxic effect under an amino acid-starving condition in head and neck squamous cancer cell lines such as CAL 27 and Detroit 562 as models of solid tumors with an upregulated autophagy in the central region owing to hypovascularity. AZM and CAM induced cell death under the amino acid-depleted (AAD) culture condition in these cell lines along with CHOP upregulation, although they showed no cytotoxicity under the complete culture medium. CHOP knockdown by siRNA in the CAL 27 cells significantly suppressed macrolide-induced cell death under the AAD culture condition. CHOP(-/-) murine embryonic fibroblast (MEF) cell lines also attenuated AZM-induced cell death compared with CHOP(+/+) MEF cell lines. Using a tet-off atg5 MEF cell line, knockout of atg5, an essential gene for autophagy, also induced cell death and CHOP in the AAD culture medium but not in the complete culture medium. This suggest that macrolide-induced cell death via CHOP induction is dependent on autophagy inhibition. The cytotoxicity of macrolide with CHOP induction was completely cancelled by the addition of amino acids in the culture medium, indicating that the cytotoxicity is due to the insufficient amino acid pool. These data suggest the possibility of using macrolides for “tumor-starving therapy”.
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spelling pubmed-51581962016-12-21 Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines Hirasawa, Kazuhiro Moriya, Shota Miyahara, Kana Kazama, Hiromi Hirota, Ayako Takemura, Jun Abe, Akihisa Inazu, Masato Hiramoto, Masaki Tsukahara, Kiyoaki Miyazawa, Keisuke PLoS One Research Article Autophagy, a self-digestive system for cytoplasmic components, is required to maintain the amino acid pool for cellular homeostasis. We previously reported that the macrolide antibiotics azithromycin (AZM) and clarithromycin (CAM) have an inhibitory effect on autophagy flux, and they potently enhance the cytocidal effect of various anticancer reagents in vitro. This suggests that macrolide antibiotics can be used as an adjuvant for cancer chemotherapy. Since cancer cells require a larger metabolic demand than normal cells because of their exuberant growth, upregulated autophagy in tumor cells has now become the target for cancer therapy. In the present study, we examined whether macrolides exhibit cytotoxic effect under an amino acid-starving condition in head and neck squamous cancer cell lines such as CAL 27 and Detroit 562 as models of solid tumors with an upregulated autophagy in the central region owing to hypovascularity. AZM and CAM induced cell death under the amino acid-depleted (AAD) culture condition in these cell lines along with CHOP upregulation, although they showed no cytotoxicity under the complete culture medium. CHOP knockdown by siRNA in the CAL 27 cells significantly suppressed macrolide-induced cell death under the AAD culture condition. CHOP(-/-) murine embryonic fibroblast (MEF) cell lines also attenuated AZM-induced cell death compared with CHOP(+/+) MEF cell lines. Using a tet-off atg5 MEF cell line, knockout of atg5, an essential gene for autophagy, also induced cell death and CHOP in the AAD culture medium but not in the complete culture medium. This suggest that macrolide-induced cell death via CHOP induction is dependent on autophagy inhibition. The cytotoxicity of macrolide with CHOP induction was completely cancelled by the addition of amino acids in the culture medium, indicating that the cytotoxicity is due to the insufficient amino acid pool. These data suggest the possibility of using macrolides for “tumor-starving therapy”. Public Library of Science 2016-12-15 /pmc/articles/PMC5158196/ /pubmed/27977675 http://dx.doi.org/10.1371/journal.pone.0164529 Text en © 2016 Hirasawa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Hirasawa, Kazuhiro
Moriya, Shota
Miyahara, Kana
Kazama, Hiromi
Hirota, Ayako
Takemura, Jun
Abe, Akihisa
Inazu, Masato
Hiramoto, Masaki
Tsukahara, Kiyoaki
Miyazawa, Keisuke
Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines
title Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines
title_full Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines
title_fullStr Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines
title_full_unstemmed Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines
title_short Macrolide Antibiotics Exhibit Cytotoxic Effect under Amino Acid-Depleted Culture Condition by Blocking Autophagy Flux in Head and Neck Squamous Cell Carcinoma Cell Lines
title_sort macrolide antibiotics exhibit cytotoxic effect under amino acid-depleted culture condition by blocking autophagy flux in head and neck squamous cell carcinoma cell lines
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5158196/
https://www.ncbi.nlm.nih.gov/pubmed/27977675
http://dx.doi.org/10.1371/journal.pone.0164529
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