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Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4

Actinomycin D (ActD) is an FDA-approved NCI oncology drug that specifically targets and downregulates stem cell transcription factors, which leads to a depletion of stem cells within the tumor bulk. Recently, our research group demonstrated the importance of IRS-4 in the development of liver cancer....

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Autores principales: Guijarro, Luis G., Sanmartin-Salinas, Patricia, Pérez-Cuevas, Eva, Toledo-Lobo, M. Val, Monserrat, Jorge, Zoullas, Sofia, Sáez, Miguel A., Álvarez-Mon, Miguel Angel, Bujan, Julia, Noguerales-Fraguas, Fernando, Arilla-Ferreiro, Eduardo, Álvarez-Mon, Melchor, Ortega, Miguel A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8472101/
https://www.ncbi.nlm.nih.gov/pubmed/34577545
http://dx.doi.org/10.3390/ph14090845
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author Guijarro, Luis G.
Sanmartin-Salinas, Patricia
Pérez-Cuevas, Eva
Toledo-Lobo, M. Val
Monserrat, Jorge
Zoullas, Sofia
Sáez, Miguel A.
Álvarez-Mon, Miguel Angel
Bujan, Julia
Noguerales-Fraguas, Fernando
Arilla-Ferreiro, Eduardo
Álvarez-Mon, Melchor
Ortega, Miguel A.
author_facet Guijarro, Luis G.
Sanmartin-Salinas, Patricia
Pérez-Cuevas, Eva
Toledo-Lobo, M. Val
Monserrat, Jorge
Zoullas, Sofia
Sáez, Miguel A.
Álvarez-Mon, Miguel Angel
Bujan, Julia
Noguerales-Fraguas, Fernando
Arilla-Ferreiro, Eduardo
Álvarez-Mon, Melchor
Ortega, Miguel A.
author_sort Guijarro, Luis G.
collection PubMed
description Actinomycin D (ActD) is an FDA-approved NCI oncology drug that specifically targets and downregulates stem cell transcription factors, which leads to a depletion of stem cells within the tumor bulk. Recently, our research group demonstrated the importance of IRS-4 in the development of liver cancer. In this study, we evaluated the protective effects of IRS-4 against ActD. For this study, three hepatocellular carcinoma cell lines (HepG2, Huh7, and Chang cells) were used to study the mechanism of actinomycin D. Most assays were carried out in the Hep G2 cell line, due to the high expression of stem cell biomarkers. We found that ActD caused HepG2 cell necroptosis characterized by DNA fragmentation, decreased mitochondrial membrane potential, cytochrome c depletion, and decreased the levels of reduced glutathione. However, we did not observe a clear increase in apoptosis markers such as annexin V presence, caspase 3 activation, or PARP fragmentation. ActD produced an activation of MAP kinases (ERK, p38, and JNK) and AKT. ActD-induced activation of AKT and MAP kinases produced an activation of the Rb-E2F cascade together with a blockage of cell cycle transitions, due to c-jun depletion. ActD led to the inhibition of pCdK1 and pH3 along with DNA fragmentation resulting in cell cycle arrest and the subsequent activation of p53-dependent cell death in the HepG2 cell line. Only JNK and AKT inhibitors were protective against the effects of ActD. N-Acetyl-L-cysteine also had a protective effect as it restored GSH levels. A likely mechanism for this is IRS-4 stimulating GCL-GSH and inhibiting the Brk-CHK1-p53 pathway. The assessment of the IRS-4 in cancer biopsies could be of interest to carry out a personalized treatment with ActD.
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spelling pubmed-84721012021-09-28 Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4 Guijarro, Luis G. Sanmartin-Salinas, Patricia Pérez-Cuevas, Eva Toledo-Lobo, M. Val Monserrat, Jorge Zoullas, Sofia Sáez, Miguel A. Álvarez-Mon, Miguel Angel Bujan, Julia Noguerales-Fraguas, Fernando Arilla-Ferreiro, Eduardo Álvarez-Mon, Melchor Ortega, Miguel A. Pharmaceuticals (Basel) Article Actinomycin D (ActD) is an FDA-approved NCI oncology drug that specifically targets and downregulates stem cell transcription factors, which leads to a depletion of stem cells within the tumor bulk. Recently, our research group demonstrated the importance of IRS-4 in the development of liver cancer. In this study, we evaluated the protective effects of IRS-4 against ActD. For this study, three hepatocellular carcinoma cell lines (HepG2, Huh7, and Chang cells) were used to study the mechanism of actinomycin D. Most assays were carried out in the Hep G2 cell line, due to the high expression of stem cell biomarkers. We found that ActD caused HepG2 cell necroptosis characterized by DNA fragmentation, decreased mitochondrial membrane potential, cytochrome c depletion, and decreased the levels of reduced glutathione. However, we did not observe a clear increase in apoptosis markers such as annexin V presence, caspase 3 activation, or PARP fragmentation. ActD produced an activation of MAP kinases (ERK, p38, and JNK) and AKT. ActD-induced activation of AKT and MAP kinases produced an activation of the Rb-E2F cascade together with a blockage of cell cycle transitions, due to c-jun depletion. ActD led to the inhibition of pCdK1 and pH3 along with DNA fragmentation resulting in cell cycle arrest and the subsequent activation of p53-dependent cell death in the HepG2 cell line. Only JNK and AKT inhibitors were protective against the effects of ActD. N-Acetyl-L-cysteine also had a protective effect as it restored GSH levels. A likely mechanism for this is IRS-4 stimulating GCL-GSH and inhibiting the Brk-CHK1-p53 pathway. The assessment of the IRS-4 in cancer biopsies could be of interest to carry out a personalized treatment with ActD. MDPI 2021-08-25 /pmc/articles/PMC8472101/ /pubmed/34577545 http://dx.doi.org/10.3390/ph14090845 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Guijarro, Luis G.
Sanmartin-Salinas, Patricia
Pérez-Cuevas, Eva
Toledo-Lobo, M. Val
Monserrat, Jorge
Zoullas, Sofia
Sáez, Miguel A.
Álvarez-Mon, Miguel Angel
Bujan, Julia
Noguerales-Fraguas, Fernando
Arilla-Ferreiro, Eduardo
Álvarez-Mon, Melchor
Ortega, Miguel A.
Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4
title Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4
title_full Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4
title_fullStr Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4
title_full_unstemmed Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4
title_short Actinomycin D Arrests Cell Cycle of Hepatocellular Carcinoma Cell Lines and Induces p53-Dependent Cell Death: A Study of the Molecular Mechanism Involved in the Protective Effect of IRS-4
title_sort actinomycin d arrests cell cycle of hepatocellular carcinoma cell lines and induces p53-dependent cell death: a study of the molecular mechanism involved in the protective effect of irs-4
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8472101/
https://www.ncbi.nlm.nih.gov/pubmed/34577545
http://dx.doi.org/10.3390/ph14090845
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