Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progression
Abstract Background The catenin beta 1 gene (CTNNB1) plays a crucial role in the malignant progression of various cancers. Recent studies have suggested that CTNNB1 hyperactivation is closely related to the occurrence and development of bladder cancer (BCa). As a member of the deubiquitinating enzym...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-09-01
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| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12967-023-04311-3 |
| _version_ | 1797556787089506304 |
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| author | Tao Liu Meng-Qi Fan Xiao-Xiao Xie Qi-Peng Shu Xue-Hua Du Lin-Zhi Qi Xiao-Dong Zhang Ming-Hui Zhang Guang Shan Run-Lei Du Shang-Ze Li |
| author_facet | Tao Liu Meng-Qi Fan Xiao-Xiao Xie Qi-Peng Shu Xue-Hua Du Lin-Zhi Qi Xiao-Dong Zhang Ming-Hui Zhang Guang Shan Run-Lei Du Shang-Ze Li |
| author_sort | Tao Liu |
| collection | DOAJ |
| description | Abstract Background The catenin beta 1 gene (CTNNB1) plays a crucial role in the malignant progression of various cancers. Recent studies have suggested that CTNNB1 hyperactivation is closely related to the occurrence and development of bladder cancer (BCa). As a member of the deubiquitinating enzyme (DUB) family, ubiquitin C-terminal hydrolase L3 (UCHL3) is abnormally expressed in various cancers. In this study, we discovered that UCHL3 is a novel oncogene in bladder cancer, suggesting it is a promising target against bladder cancer. Methods We utilized CRISPR‒Cas9 technology to construct cell lines with UCHL3 stably overexpressed or knocked out. The successful overexpression or knockout of UCHL3 was determined using Western blotting. Then, we performed CCK-8, colony formation, soft agar and Transwell migration assays to determine the impact of the UCHL3 gene on cell phenotype. RNA-seq was performed with UCHL3-depleted T24 cells (established via CRISPR–Cas9-mediated genomic editing). We analyzed differences in WNT pathway gene expression in wild-type and UCHL3-deficient T24 cell lines using a heatmap and by gene set enrichment analysis (GSEA). Then, we validated the effect of UCHL3 on the Wnt pathway using a dual fluorescence reporter. We then analyzed the underlying mechanisms involved using Western blots, co-IP, and immunofluorescence results. We also conducted nude mouse tumor formation experiments. Moreover, conditional UCHL3-knockout mice and bladder cancer model mice were established for research. Results We found that the overexpression of UCHL3 boosted bladder cancer cell proliferation, invasion and migration, while the depletion of UCHL3 in bladder cancer cells delayed tumor tumorigenesis in vitro and in vivo. UCHL3 was highly associated with the Wnt signaling pathway and triggered the activation of the Wnt signaling pathway, which showed that its functions depend on its deubiquitination activity. Notably, Uchl3-deficient mice were less susceptible to bladder tumorigenesis. Additionally, UCHL3 was highly expressed in bladder cancer cells and associated with indicators of advanced clinicopathology. Conclusion In summary, we found that UCHL3 is amplified in bladder cancer and functions as a tumor promoter that enhances proliferation and migration of tumor cells in vitro and bladder tumorigenesis and progression in vivo. Furthermore, we revealed that UCHL3 stabilizes CTNNB1 expression, resulting in the activation of the oncogenic Wnt signaling pathway. Therefore, our findings strongly suggest that UCHL3 is a promising therapeutic target for bladder cancer. |
| first_indexed | 2024-03-10T17:07:54Z |
| format | Article |
| id | doaj.art-77b0d8d306b94929b2993eeac723798f |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-03-10T17:07:54Z |
| publishDate | 2023-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-77b0d8d306b94929b2993eeac723798f2023-11-20T10:45:18ZengBMCJournal of Translational Medicine1479-58762023-09-0121111610.1186/s12967-023-04311-3Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progressionTao Liu0Meng-Qi Fan1Xiao-Xiao Xie2Qi-Peng Shu3Xue-Hua Du4Lin-Zhi Qi5Xiao-Dong Zhang6Ming-Hui Zhang7Guang Shan8Run-Lei Du9Shang-Ze Li10Department of Urology, Zhongnan Hospital of Wuhan UniversityHubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan UniversitySchool of Medicine, Chongqing UniversityHubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan UniversityHubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan UniversitySchool of Medicine, Chongqing UniversityHubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan UniversityDepartment of Obstetrics and Gynecology, The First Affiliated Hospital of Zhengzhou UniversityDepartment of Urology, Renmin Hospital of Wuhan UniversityHubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan UniversitySchool of Medicine, Chongqing UniversityAbstract Background The catenin beta 1 gene (CTNNB1) plays a crucial role in the malignant progression of various cancers. Recent studies have suggested that CTNNB1 hyperactivation is closely related to the occurrence and development of bladder cancer (BCa). As a member of the deubiquitinating enzyme (DUB) family, ubiquitin C-terminal hydrolase L3 (UCHL3) is abnormally expressed in various cancers. In this study, we discovered that UCHL3 is a novel oncogene in bladder cancer, suggesting it is a promising target against bladder cancer. Methods We utilized CRISPR‒Cas9 technology to construct cell lines with UCHL3 stably overexpressed or knocked out. The successful overexpression or knockout of UCHL3 was determined using Western blotting. Then, we performed CCK-8, colony formation, soft agar and Transwell migration assays to determine the impact of the UCHL3 gene on cell phenotype. RNA-seq was performed with UCHL3-depleted T24 cells (established via CRISPR–Cas9-mediated genomic editing). We analyzed differences in WNT pathway gene expression in wild-type and UCHL3-deficient T24 cell lines using a heatmap and by gene set enrichment analysis (GSEA). Then, we validated the effect of UCHL3 on the Wnt pathway using a dual fluorescence reporter. We then analyzed the underlying mechanisms involved using Western blots, co-IP, and immunofluorescence results. We also conducted nude mouse tumor formation experiments. Moreover, conditional UCHL3-knockout mice and bladder cancer model mice were established for research. Results We found that the overexpression of UCHL3 boosted bladder cancer cell proliferation, invasion and migration, while the depletion of UCHL3 in bladder cancer cells delayed tumor tumorigenesis in vitro and in vivo. UCHL3 was highly associated with the Wnt signaling pathway and triggered the activation of the Wnt signaling pathway, which showed that its functions depend on its deubiquitination activity. Notably, Uchl3-deficient mice were less susceptible to bladder tumorigenesis. Additionally, UCHL3 was highly expressed in bladder cancer cells and associated with indicators of advanced clinicopathology. Conclusion In summary, we found that UCHL3 is amplified in bladder cancer and functions as a tumor promoter that enhances proliferation and migration of tumor cells in vitro and bladder tumorigenesis and progression in vivo. Furthermore, we revealed that UCHL3 stabilizes CTNNB1 expression, resulting in the activation of the oncogenic Wnt signaling pathway. Therefore, our findings strongly suggest that UCHL3 is a promising therapeutic target for bladder cancer.https://doi.org/10.1186/s12967-023-04311-3UCHL3Bladder cancerWnt signaling pathwayCTNNB1 |
| spellingShingle | Tao Liu Meng-Qi Fan Xiao-Xiao Xie Qi-Peng Shu Xue-Hua Du Lin-Zhi Qi Xiao-Dong Zhang Ming-Hui Zhang Guang Shan Run-Lei Du Shang-Ze Li Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progression Journal of Translational Medicine UCHL3 Bladder cancer Wnt signaling pathway CTNNB1 |
| title | Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progression |
| title_full | Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progression |
| title_fullStr | Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progression |
| title_full_unstemmed | Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progression |
| title_short | Activation of CTNNB1 by deubiquitinase UCHL3-mediated stabilization facilitates bladder cancer progression |
| title_sort | activation of ctnnb1 by deubiquitinase uchl3 mediated stabilization facilitates bladder cancer progression |
| topic | UCHL3 Bladder cancer Wnt signaling pathway CTNNB1 |
| url | https://doi.org/10.1186/s12967-023-04311-3 |
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