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AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells

AZD1775 is a small molecule WEE1 inhibitor used in combination with DNA-damaging agents to cause premature mitosis and cell death in p53-mutated cancer cells. Here we sought to determine the mechanism of action of AZD1775 in combination with chemotherapeutic agents in light of recent findings that A...

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Autores principales: Webster, Peter John, Littlejohns, Anna Tiffany, Gaunt, Hannah Jane, Prasad, K. Raj, Beech, David John, Burke, Dermot Anthony
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5736347/
https://www.ncbi.nlm.nih.gov/pubmed/28296564
http://dx.doi.org/10.1080/15384101.2017.1301329
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author Webster, Peter John
Littlejohns, Anna Tiffany
Gaunt, Hannah Jane
Prasad, K. Raj
Beech, David John
Burke, Dermot Anthony
author_facet Webster, Peter John
Littlejohns, Anna Tiffany
Gaunt, Hannah Jane
Prasad, K. Raj
Beech, David John
Burke, Dermot Anthony
author_sort Webster, Peter John
collection PubMed
description AZD1775 is a small molecule WEE1 inhibitor used in combination with DNA-damaging agents to cause premature mitosis and cell death in p53-mutated cancer cells. Here we sought to determine the mechanism of action of AZD1775 in combination with chemotherapeutic agents in light of recent findings that AZD1775 can cause double-stranded DNA (DS-DNA) breaks. AZD1775 significantly improved the cytotoxicity of 5-FU in a p53-mutated colorectal cancer cell line (HT29 cells), decreasing the IC(50) from 9.3 μM to 3.5 μM. Flow cytometry showed a significant increase in the mitotic marker pHH3 (3.4% vs. 56.2%) and DS-DNA break marker γH2AX (5.1% vs. 50.7%) for combination therapy compared with 5-FU alone. Combination therapy also increased the amount of caspase-3 dependent apoptosis compared with 5-FU alone (4% vs. 13%). The addition of exogenous nucleosides to combination therapy significantly rescued the increased DS-DNA breaks and caspase-3 dependent apoptosis almost to the levels of 5-FU monotherapy. In conclusion, AZD1775 enhances 5-FU cytotoxicity through increased DS-DNA breaks, not premature mitosis, in p53-mutated colorectal cancer cells. This finding is important for designers of future clinical trials when considering the optimal timing and duration of AZD1775 treatment.
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spelling pubmed-57363472017-12-22 AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells Webster, Peter John Littlejohns, Anna Tiffany Gaunt, Hannah Jane Prasad, K. Raj Beech, David John Burke, Dermot Anthony Cell Cycle Report AZD1775 is a small molecule WEE1 inhibitor used in combination with DNA-damaging agents to cause premature mitosis and cell death in p53-mutated cancer cells. Here we sought to determine the mechanism of action of AZD1775 in combination with chemotherapeutic agents in light of recent findings that AZD1775 can cause double-stranded DNA (DS-DNA) breaks. AZD1775 significantly improved the cytotoxicity of 5-FU in a p53-mutated colorectal cancer cell line (HT29 cells), decreasing the IC(50) from 9.3 μM to 3.5 μM. Flow cytometry showed a significant increase in the mitotic marker pHH3 (3.4% vs. 56.2%) and DS-DNA break marker γH2AX (5.1% vs. 50.7%) for combination therapy compared with 5-FU alone. Combination therapy also increased the amount of caspase-3 dependent apoptosis compared with 5-FU alone (4% vs. 13%). The addition of exogenous nucleosides to combination therapy significantly rescued the increased DS-DNA breaks and caspase-3 dependent apoptosis almost to the levels of 5-FU monotherapy. In conclusion, AZD1775 enhances 5-FU cytotoxicity through increased DS-DNA breaks, not premature mitosis, in p53-mutated colorectal cancer cells. This finding is important for designers of future clinical trials when considering the optimal timing and duration of AZD1775 treatment. Taylor & Francis 2017-11-09 /pmc/articles/PMC5736347/ /pubmed/28296564 http://dx.doi.org/10.1080/15384101.2017.1301329 Text en © 2017 Peter John Webster, Anna Tiffany Littlejohns, Hannah Jane Gaunt, K. Raj Prasad, David John Beech, and Dermot Anthony Burke. Published with license by Taylor & Francis. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.
spellingShingle Report
Webster, Peter John
Littlejohns, Anna Tiffany
Gaunt, Hannah Jane
Prasad, K. Raj
Beech, David John
Burke, Dermot Anthony
AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
title AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
title_full AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
title_fullStr AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
title_full_unstemmed AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
title_short AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
title_sort azd1775 induces toxicity through double-stranded dna breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5736347/
https://www.ncbi.nlm.nih.gov/pubmed/28296564
http://dx.doi.org/10.1080/15384101.2017.1301329
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