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A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299

Autophagy inhibition through small-molecule inhibitors is one of the approaches to increase the efficiency of radiotherapy in oncological patients. A new inhibitor—Lys05—with the potential to accumulate within lysosomes and to block autophagy was discovered a few years ago. Several studies have addr...

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Autores principales: Cechakova, Lucie, Ondrej, Martin, Pavlik, Vojtech, Jost, Petr, Cizkova, Dana, Bezrouk, Ales, Pejchal, Jaroslav, Amaravadi, Ravi K., Winkler, Jeffrey D., Tichy, Ales
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6928878/
https://www.ncbi.nlm.nih.gov/pubmed/31771188
http://dx.doi.org/10.3390/ijms20235881
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author Cechakova, Lucie
Ondrej, Martin
Pavlik, Vojtech
Jost, Petr
Cizkova, Dana
Bezrouk, Ales
Pejchal, Jaroslav
Amaravadi, Ravi K.
Winkler, Jeffrey D.
Tichy, Ales
author_facet Cechakova, Lucie
Ondrej, Martin
Pavlik, Vojtech
Jost, Petr
Cizkova, Dana
Bezrouk, Ales
Pejchal, Jaroslav
Amaravadi, Ravi K.
Winkler, Jeffrey D.
Tichy, Ales
author_sort Cechakova, Lucie
collection PubMed
description Autophagy inhibition through small-molecule inhibitors is one of the approaches to increase the efficiency of radiotherapy in oncological patients. A new inhibitor—Lys05—with the potential to accumulate within lysosomes and to block autophagy was discovered a few years ago. Several studies have addressed its chemosensitizing effects but nothing is known about its impact in the context of ionizing radiation (IR). To describe its role in radiosensitization, we employed radioresistant human non-small cell lung carcinoma cells (H1299, p53-negative). Combined treatment of H1299 cells by Lys05 together with IR decreased cell survival in the clonogenic assay and real-time monitoring of cell growth more than either Lys05 or IR alone. Immunodetection of LC3 and p62/SQSTM1 indicated that autophagy was inhibited, which correlated with increased SQSTM1 and decreased BNIP3 gene expression determined by qRT-PCR. Fluorescence microscopy and flow cytometry uncovered an accumulation of lysosomes. Similarly, transmission electron microscopy demonstrated the accumulation of autophagosomes confirming the ability of Lys05 to potentiate autophagy inhibition in H1299 cells. We report here for the first time that Lys05 could be utilized in combination with IR as a promising future strategy in the eradication of lung cancer cells.
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spelling pubmed-69288782019-12-26 A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299 Cechakova, Lucie Ondrej, Martin Pavlik, Vojtech Jost, Petr Cizkova, Dana Bezrouk, Ales Pejchal, Jaroslav Amaravadi, Ravi K. Winkler, Jeffrey D. Tichy, Ales Int J Mol Sci Article Autophagy inhibition through small-molecule inhibitors is one of the approaches to increase the efficiency of radiotherapy in oncological patients. A new inhibitor—Lys05—with the potential to accumulate within lysosomes and to block autophagy was discovered a few years ago. Several studies have addressed its chemosensitizing effects but nothing is known about its impact in the context of ionizing radiation (IR). To describe its role in radiosensitization, we employed radioresistant human non-small cell lung carcinoma cells (H1299, p53-negative). Combined treatment of H1299 cells by Lys05 together with IR decreased cell survival in the clonogenic assay and real-time monitoring of cell growth more than either Lys05 or IR alone. Immunodetection of LC3 and p62/SQSTM1 indicated that autophagy was inhibited, which correlated with increased SQSTM1 and decreased BNIP3 gene expression determined by qRT-PCR. Fluorescence microscopy and flow cytometry uncovered an accumulation of lysosomes. Similarly, transmission electron microscopy demonstrated the accumulation of autophagosomes confirming the ability of Lys05 to potentiate autophagy inhibition in H1299 cells. We report here for the first time that Lys05 could be utilized in combination with IR as a promising future strategy in the eradication of lung cancer cells. MDPI 2019-11-23 /pmc/articles/PMC6928878/ /pubmed/31771188 http://dx.doi.org/10.3390/ijms20235881 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cechakova, Lucie
Ondrej, Martin
Pavlik, Vojtech
Jost, Petr
Cizkova, Dana
Bezrouk, Ales
Pejchal, Jaroslav
Amaravadi, Ravi K.
Winkler, Jeffrey D.
Tichy, Ales
A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299
title A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299
title_full A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299
title_fullStr A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299
title_full_unstemmed A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299
title_short A Potent Autophagy Inhibitor (Lys05) Enhances the Impact of Ionizing Radiation on Human Lung Cancer Cells H1299
title_sort potent autophagy inhibitor (lys05) enhances the impact of ionizing radiation on human lung cancer cells h1299
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6928878/
https://www.ncbi.nlm.nih.gov/pubmed/31771188
http://dx.doi.org/10.3390/ijms20235881
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