Bufalin Inhibits Tumorigenesis, Stemness, and Epithelial–Mesenchymal Transition in Colorectal Cancer through a C-Kit/Slug Signaling Axis

• Published in: International journal of molecular sciences Vol. 23; no. 21; p. 13354
• Main Authors: Ding, Ling, Yang, Yuning, Lu, Qin, Qu, Dongfeng, Chandrakesan, Parthasarathy, Feng, Hailan, Chen, Hong, Chen, Xuzheng, Liao, Zhuhui, Du, Jian, Cao, Zhiyun, Weygant, Nathaniel
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01-11-2022; MDPI
• Subjects: Aldehyde dehydrogenase; Aldehydes; Apoptosis; C-Kit; c-Kit protein; c-Myc protein; Cancer; cancer stem cells; CD44 antigen; Colorectal cancer; Colorectal carcinoma; Drug resistance; E-cadherin; Feedback loops; Gene expression; Heterogeneity; Kinases; Markers; Medical prognosis; Mesenchyme; Metastasis; Morbidity; Myc protein; N-Cadherin; Network analysis; Nuclear transport; Organoids; Ovarian cancer; patient-derived organoids; personalized therapy; Pluripotency; Prostate cancer; Slug; Stem cells; Transcription factors; Tumorigenesis; Tumors; β-Catenin
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms232113354

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.