Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loop
Introduction: Tumor-associated calcium signal transducer 2 (Trop2) has been used as a transport gate for cytotoxic agents into cells in antibody–drug conjugate designs because of its expression in a wide range of solid tumors. However, the specific role of Trop2 itself in breast cancer progression r...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-04-01
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| Series: | Journal of Advanced Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2090123223001509 |
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| author | Wenjuan Tang Yu Hu Kaihui Tu Zhengyan Gong Man Zhu Tianfeng Yang Ammar Sarwar Bingling Dai Dongdong Zhang Yingzhuan Zhan Yanmin Zhang |
| author_facet | Wenjuan Tang Yu Hu Kaihui Tu Zhengyan Gong Man Zhu Tianfeng Yang Ammar Sarwar Bingling Dai Dongdong Zhang Yingzhuan Zhan Yanmin Zhang |
| author_sort | Wenjuan Tang |
| collection | DOAJ |
| description | Introduction: Tumor-associated calcium signal transducer 2 (Trop2) has been used as a transport gate for cytotoxic agents into cells in antibody–drug conjugate designs because of its expression in a wide range of solid tumors. However, the specific role of Trop2 itself in breast cancer progression remains unclear and small molecules targeting Trop2 have not yet been reported. Objectives: To screen small molecules targeting Trop2, and to reveal its pharmacological effects and the molecular mechanisms of action. Methods: Small molecule targeting Trop2 was identified by cell membrane chromatography, and validated by cellular thermal shift assay and point mutation analyses. We investigated the pharmacological effects of Trop2 inhibitor using RNA-seq, human foreskin fibroblast (HFF)-derived extracellular matrix (ECM), Matrigel drop invasion assays, colony-forming assay, xenograft tumor model, 4T1 orthotopic metastasis model and 4T1 experimental metastasis model. The molecular mechanism was determined using immunoprecipitation, mass spectrometry, immunofluorescence, immunohistochemistry and Western blotting. Results: Here we identified Bruceine D (BD) as the inhibitor of Trop2, and demonstrated anti-metastasis effects of BD in breast cancer. Notably, Lys307 and Glu310 residues of Trop2 acted as critical sites for BD binding. Mechanistically, BD suppressed Trop2-induced cancer metastasis by blocking the formation of Trop2/β-catenin positive loop, in which the Trop2/β-catenin complex prevented β-catenin from being degraded via the ubiquitin-proteosome pathway. Destabilized β-catenin caused by BD reduced nucleus translocation, leading to the reduction of transcription of Trop2, the reversal of epithelial-mesenchymal transition (EMT) process, and the inhibition of ECM remodeling, further inhibiting cancer metastasis. Additionally, the inhibitory effects of BD on lung metastatic colonization and the beneficial effects of BD on prolongation of survival were validated in 4T1 experimental metastasis model. Conclusions: These results support the tumor-promoting role of Trop2 in breast cancer by stabilizing β-catenin in Trop2/β-catenin positive loop, and suggest Bruceine D as a promising candidate for Trop2 inhibition. |
| first_indexed | 2024-04-24T19:48:36Z |
| format | Article |
| id | doaj.art-877e82d55c484d0aadcaadee5090206b |
| institution | Directory Open Access Journal |
| issn | 2090-1232 |
| language | English |
| last_indexed | 2024-04-24T19:48:36Z |
| publishDate | 2024-04-01 |
| publisher | Elsevier |
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| series | Journal of Advanced Research |
| spelling | doaj.art-877e82d55c484d0aadcaadee5090206b2024-03-25T04:17:27ZengElsevierJournal of Advanced Research2090-12322024-04-0158193210Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loopWenjuan Tang0Yu Hu1Kaihui Tu2Zhengyan Gong3Man Zhu4Tianfeng Yang5Ammar Sarwar6Bingling Dai7Dongdong Zhang8Yingzhuan Zhan9Yanmin Zhang10School of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaSchool of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaCorresponding authors at: School of Pharmacy, Health Science Center, Xi’an Jiaotong University, No. 76, Yanta Weststreet, #54, Xi’an, Shaanxi Province 710061, China.; School of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaCorresponding authors at: School of Pharmacy, Health Science Center, Xi’an Jiaotong University, No. 76, Yanta Weststreet, #54, Xi’an, Shaanxi Province 710061, China.; School of Pharmacy, Health Science Center, Xi’an Jiaotong University, Xi’an 710061, China; State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi’an 710061, ChinaIntroduction: Tumor-associated calcium signal transducer 2 (Trop2) has been used as a transport gate for cytotoxic agents into cells in antibody–drug conjugate designs because of its expression in a wide range of solid tumors. However, the specific role of Trop2 itself in breast cancer progression remains unclear and small molecules targeting Trop2 have not yet been reported. Objectives: To screen small molecules targeting Trop2, and to reveal its pharmacological effects and the molecular mechanisms of action. Methods: Small molecule targeting Trop2 was identified by cell membrane chromatography, and validated by cellular thermal shift assay and point mutation analyses. We investigated the pharmacological effects of Trop2 inhibitor using RNA-seq, human foreskin fibroblast (HFF)-derived extracellular matrix (ECM), Matrigel drop invasion assays, colony-forming assay, xenograft tumor model, 4T1 orthotopic metastasis model and 4T1 experimental metastasis model. The molecular mechanism was determined using immunoprecipitation, mass spectrometry, immunofluorescence, immunohistochemistry and Western blotting. Results: Here we identified Bruceine D (BD) as the inhibitor of Trop2, and demonstrated anti-metastasis effects of BD in breast cancer. Notably, Lys307 and Glu310 residues of Trop2 acted as critical sites for BD binding. Mechanistically, BD suppressed Trop2-induced cancer metastasis by blocking the formation of Trop2/β-catenin positive loop, in which the Trop2/β-catenin complex prevented β-catenin from being degraded via the ubiquitin-proteosome pathway. Destabilized β-catenin caused by BD reduced nucleus translocation, leading to the reduction of transcription of Trop2, the reversal of epithelial-mesenchymal transition (EMT) process, and the inhibition of ECM remodeling, further inhibiting cancer metastasis. Additionally, the inhibitory effects of BD on lung metastatic colonization and the beneficial effects of BD on prolongation of survival were validated in 4T1 experimental metastasis model. Conclusions: These results support the tumor-promoting role of Trop2 in breast cancer by stabilizing β-catenin in Trop2/β-catenin positive loop, and suggest Bruceine D as a promising candidate for Trop2 inhibition.http://www.sciencedirect.com/science/article/pii/S2090123223001509Trop2β-cateninBreast cancerMetastasisLung colonization |
| spellingShingle | Wenjuan Tang Yu Hu Kaihui Tu Zhengyan Gong Man Zhu Tianfeng Yang Ammar Sarwar Bingling Dai Dongdong Zhang Yingzhuan Zhan Yanmin Zhang Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loop Journal of Advanced Research Trop2 β-catenin Breast cancer Metastasis Lung colonization |
| title | Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loop |
| title_full | Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loop |
| title_fullStr | Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loop |
| title_full_unstemmed | Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loop |
| title_short | Targeting Trop2 by Bruceine D suppresses breast cancer metastasis by blocking Trop2/β-catenin positive feedback loop |
| title_sort | targeting trop2 by bruceine d suppresses breast cancer metastasis by blocking trop2 β catenin positive feedback loop |
| topic | Trop2 β-catenin Breast cancer Metastasis Lung colonization |
| url | http://www.sciencedirect.com/science/article/pii/S2090123223001509 |
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