Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis
Colorectal cancer (CRC) is a major source of morbidity and mortality, characterized by intratumoral heterogeneity and the presence of cancer stem cells (CSCs). Bufalin has potent activity against many tumors, but studies of its effect on CRC stemness are limited. We explored bufalin’s function and m...
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2022-11-01
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author | Ling Ding Yuning Yang Qin Lu Dongfeng Qu Parthasarathy Chandrakesan Hailan Feng Hong Chen Xuzheng Chen Zhuhui Liao Jian Du Zhiyun Cao Nathaniel Weygant |
author_facet | Ling Ding Yuning Yang Qin Lu Dongfeng Qu Parthasarathy Chandrakesan Hailan Feng Hong Chen Xuzheng Chen Zhuhui Liao Jian Du Zhiyun Cao Nathaniel Weygant |
author_sort | Ling Ding |
collection | DOAJ |
description | Colorectal cancer (CRC) is a major source of morbidity and mortality, characterized by intratumoral heterogeneity and the presence of cancer stem cells (CSCs). Bufalin has potent activity against many tumors, but studies of its effect on CRC stemness are limited. We explored bufalin’s function and mechanism using CRC patient-derived organoids (PDOs) and cell lines. In CRC cells, bufalin prevented nuclear translocation of β-catenin and down-regulated CSC markers (CD44, CD133, LGR5), pluripotency factors, and epithelial–mesenchymal transition (EMT) markers (N-Cadherin, Slug, ZEB1). Functionally, bufalin inhibited CRC spheroid formation, aldehyde dehydrogenase activity, migration, and invasion. Network analysis identified a C-Kit/Slug signaling axis accounting for bufalin’s anti-stemness activity. Bufalin treatment significantly downregulated C-Kit, as predicted. Furthermore, overexpression of C-Kit induced Slug expression, spheroid formation, and bufalin resistance. Similarly, overexpression of Slug resulted in increased expression of C-Kit and identical functional effects, demonstrating a pro-stemness feedback loop. For further study, we established PDOs from diagnostic colonoscopy. Bufalin differentially inhibited PDO growth and proliferation, induced apoptosis, restored E-cadherin, and downregulated CSC markers CD133 and C-Myc, dependent on C-Kit/Slug. These findings suggest that the C-Kit/Slug axis plays a pivotal role in regulating CRC stemness, and reveal that targeting this axis can inhibit CRC growth and progression. |
first_indexed | 2024-03-09T19:00:02Z |
format | Article |
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issn | 1661-6596 1422-0067 |
language | English |
last_indexed | 2024-03-09T19:00:02Z |
publishDate | 2022-11-01 |
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series | International Journal of Molecular Sciences |
spelling | doaj.art-d4766299aea047c692350c0020c8fd2c2023-11-24T05:06:36ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-11-0123211335410.3390/ijms232113354Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling AxisLing Ding0Yuning Yang1Qin Lu2Dongfeng Qu3Parthasarathy Chandrakesan4Hailan Feng5Hong Chen6Xuzheng Chen7Zhuhui Liao8Jian Du9Zhiyun Cao10Nathaniel Weygant11Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaDepartment of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USADepartment of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USAAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaAcademy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, ChinaColorectal cancer (CRC) is a major source of morbidity and mortality, characterized by intratumoral heterogeneity and the presence of cancer stem cells (CSCs). Bufalin has potent activity against many tumors, but studies of its effect on CRC stemness are limited. We explored bufalin’s function and mechanism using CRC patient-derived organoids (PDOs) and cell lines. In CRC cells, bufalin prevented nuclear translocation of β-catenin and down-regulated CSC markers (CD44, CD133, LGR5), pluripotency factors, and epithelial–mesenchymal transition (EMT) markers (N-Cadherin, Slug, ZEB1). Functionally, bufalin inhibited CRC spheroid formation, aldehyde dehydrogenase activity, migration, and invasion. Network analysis identified a C-Kit/Slug signaling axis accounting for bufalin’s anti-stemness activity. Bufalin treatment significantly downregulated C-Kit, as predicted. Furthermore, overexpression of C-Kit induced Slug expression, spheroid formation, and bufalin resistance. Similarly, overexpression of Slug resulted in increased expression of C-Kit and identical functional effects, demonstrating a pro-stemness feedback loop. For further study, we established PDOs from diagnostic colonoscopy. Bufalin differentially inhibited PDO growth and proliferation, induced apoptosis, restored E-cadherin, and downregulated CSC markers CD133 and C-Myc, dependent on C-Kit/Slug. These findings suggest that the C-Kit/Slug axis plays a pivotal role in regulating CRC stemness, and reveal that targeting this axis can inhibit CRC growth and progression.https://www.mdpi.com/1422-0067/23/21/13354patient-derived organoidspersonalized therapycolorectal cancercancer stem cellsC-KitSlug |
spellingShingle | Ling Ding Yuning Yang Qin Lu Dongfeng Qu Parthasarathy Chandrakesan Hailan Feng Hong Chen Xuzheng Chen Zhuhui Liao Jian Du Zhiyun Cao Nathaniel Weygant Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis International Journal of Molecular Sciences patient-derived organoids personalized therapy colorectal cancer cancer stem cells C-Kit Slug |
title | Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis |
title_full | Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis |
title_fullStr | Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis |
title_full_unstemmed | Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis |
title_short | Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis |
title_sort | bufalin inhibits tumorigenesis stemness and epithelial mesenchymal transition in colorectal cancer through a c kit slug signaling axis |
topic | patient-derived organoids personalized therapy colorectal cancer cancer stem cells C-Kit Slug |
url | https://www.mdpi.com/1422-0067/23/21/13354 |
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