Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation
Abstract 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 p...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-01-01
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| Series: | Molecular Medicine |
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| Online Access: | https://doi.org/10.1186/s10020-023-00608-7 |
| _version_ | 1797945827895803904 |
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| author | Mayao Luo Yifan Zhang Zhuofan Xu Shidong Lv Qiang Wei Qiang Dang |
| author_facet | Mayao Luo Yifan Zhang Zhuofan Xu Shidong Lv Qiang Wei Qiang Dang |
| author_sort | Mayao Luo |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-04-10T21:02:26Z |
| format | Article |
| id | doaj.art-b7055715f5814db38d9defd953b20219 |
| institution | Directory Open Access Journal |
| issn | 1528-3658 |
| language | English |
| last_indexed | 2024-04-10T21:02:26Z |
| publishDate | 2023-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Medicine |
| spelling | doaj.art-b7055715f5814db38d9defd953b202192023-01-22T12:17:04ZengBMCMolecular Medicine1528-36582023-01-0129111110.1186/s10020-023-00608-7Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutationMayao Luo0Yifan Zhang1Zhuofan Xu2Shidong Lv3Qiang Wei4Qiang Dang5Department of Urology, Nanfang Hospital, Southern Medical UniversityDepartment of Urology, Nanfang Hospital, Southern Medical UniversityDepartment of Urology, Nanfang Hospital, Southern Medical UniversityDepartment of Urology, Nanfang Hospital, Southern Medical UniversityDepartment of Urology, Nanfang Hospital, Southern Medical UniversityDepartment of Urology, Nanfang Hospital, Southern Medical UniversityAbstract 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.https://doi.org/10.1186/s10020-023-00608-7EP300Bladder cancerMissense mutationDriver mutation |
| spellingShingle | Mayao Luo Yifan Zhang Zhuofan Xu Shidong Lv Qiang Wei Qiang Dang Experimental analysis of bladder cancer-associated mutations in EP300 identifies EP300-R1627W as a driver mutation Molecular Medicine EP300 Bladder cancer Missense mutation 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 | EP300 Bladder cancer Missense mutation Driver mutation |
| url | https://doi.org/10.1186/s10020-023-00608-7 |
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