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Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation

BACKGROUND: Bladder cancer (BCa) is the most common malignant tumor of the urinary system, with transitional cell carcinoma (TCC) being the predominant type. EP300 encodes a lysine acetyltransferase that regulates a large subset of genes by acetylating histones and non-histone proteins. We previousl...

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Autores principales: Luo, Mayao, Zhang, Yifan, Xu, Zhuofan, Lv, Shidong, Wei, Qiang, Dang, Qiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9843983/
https://www.ncbi.nlm.nih.gov/pubmed/36647005
http://dx.doi.org/10.1186/s10020-023-00608-7
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author Luo, Mayao
Zhang, Yifan
Xu, Zhuofan
Lv, Shidong
Wei, Qiang
Dang, Qiang
author_facet Luo, Mayao
Zhang, Yifan
Xu, Zhuofan
Lv, Shidong
Wei, Qiang
Dang, Qiang
author_sort Luo, Mayao
collection PubMed
description BACKGROUND: Bladder cancer (BCa) is the most common malignant tumor of the urinary system, with transitional cell carcinoma (TCC) being the predominant type. EP300 encodes a lysine acetyltransferase that regulates a large subset of genes by acetylating histones and non-histone proteins. We previously identified several bladder cancer-associated mutations in EP300 using high-throughput sequencing; however, the functional consequences of these mutations remain unclear. METHODS: Bladder cancer cells T24 and TCC-SUP were infected with shEP300 lentiviruses to generate stable EP300 knockdown cell lines. The expression levels of EP300, p16 and p21 were detected by real-time PCR and western blots. The transcriptional activity of p16 and p21 were detected by dual luciferase assay. Cell proliferation assay, flow cytometric analyses of cell cycle, invasion assay and xenograft tumor model were used to measure the effect of EP300-R1627W mutation in bladder cancer. Immunoprecipitation was used to explore the relationship between EP300-R1627W mutation and p53. Structural analysis was used to detect the structure of EP300-R1627W protein compared to EP300-wt protein. RESULTS: we screened the mutations of EP300 and found that the EP300-R1627W mutation significantly impairs EP300 transactivation activity. Notably, we demonstrated that the R1627W mutation impairs EP300 acetyltransferase activity, potentially by interfering with substrate binding. Finally, we show that EP300-R1627W is more aggressive in growth and invasion in vitro and in vivo compared to cells expressing EP300-wt. We also found that the EP300-R1627W mutation occurs frequently in seven different types of cancers. CONCLUSION: In summary, our work defines a driver role of EP300-R1627W in bladder cancer development and progression. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s10020-023-00608-7.
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spelling pubmed-98439832023-01-18 Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation Luo, Mayao Zhang, Yifan Xu, Zhuofan Lv, Shidong Wei, Qiang Dang, Qiang Mol Med Research Article BACKGROUND: Bladder cancer (BCa) is the most common malignant tumor of the urinary system, with transitional cell carcinoma (TCC) being the predominant type. EP300 encodes a lysine acetyltransferase that regulates a large subset of genes by acetylating histones and non-histone proteins. We previously identified several bladder cancer-associated mutations in EP300 using high-throughput sequencing; however, the functional consequences of these mutations remain unclear. METHODS: Bladder cancer cells T24 and TCC-SUP were infected with shEP300 lentiviruses to generate stable EP300 knockdown cell lines. The expression levels of EP300, p16 and p21 were detected by real-time PCR and western blots. The transcriptional activity of p16 and p21 were detected by dual luciferase assay. Cell proliferation assay, flow cytometric analyses of cell cycle, invasion assay and xenograft tumor model were used to measure the effect of EP300-R1627W mutation in bladder cancer. Immunoprecipitation was used to explore the relationship between EP300-R1627W mutation and p53. Structural analysis was used to detect the structure of EP300-R1627W protein compared to EP300-wt protein. RESULTS: we screened the mutations of EP300 and found that the EP300-R1627W mutation significantly impairs EP300 transactivation activity. Notably, we demonstrated that the R1627W mutation impairs EP300 acetyltransferase activity, potentially by interfering with substrate binding. Finally, we show that EP300-R1627W is more aggressive in growth and invasion in vitro and in vivo compared to cells expressing EP300-wt. We also found that the EP300-R1627W mutation occurs frequently in seven different types of cancers. CONCLUSION: In summary, our work defines a driver role of EP300-R1627W in bladder cancer development and progression. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s10020-023-00608-7. BioMed Central 2023-01-16 /pmc/articles/PMC9843983/ /pubmed/36647005 http://dx.doi.org/10.1186/s10020-023-00608-7 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Luo, Mayao
Zhang, Yifan
Xu, Zhuofan
Lv, Shidong
Wei, Qiang
Dang, Qiang
Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation
title Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation
title_full Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation
title_fullStr Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation
title_full_unstemmed Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation
title_short Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation
title_sort experimental analysis of bladder cancer-associated mutations in ep300 identifies ep300-r1627w as a driver mutation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9843983/
https://www.ncbi.nlm.nih.gov/pubmed/36647005
http://dx.doi.org/10.1186/s10020-023-00608-7
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