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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Main Authors: Ling Ding, Yuning Yang, Qin Lu, Dongfeng Qu, Parthasarathy Chandrakesan, Hailan Feng, Hong Chen, Xuzheng Chen, Zhuhui Liao, Jian Du, Zhiyun Cao, Nathaniel Weygant
Format: Article
Language:English
Published: MDPI AG 2022-11-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/23/21/13354
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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.
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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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