Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Feb 26, 2024; 16(2): 207-227
Published online Feb 26, 2024. doi: 10.4252/wjsc.v16.i2.207
VX-509 attenuates the stemness characteristics of colorectal cancer stem-like cells by regulating the epithelial-mesenchymal transition through Nodal/Smad2/3 signaling
Yun Yuan, Xu-Fan Zhang, Yu-Chen Li, Hong-Qing Chen, Tian Wen, Jia-Lian Zheng, Zi-Yi Zhao, Qiong-Ying Hu
Yun Yuan, Yu-Chen Li, Hong-Qing Chen, Tian Wen, Qiong-Ying Hu, Department of Laboratory Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu 610072, Sichuan Province, China
Xu-Fan Zhang, Department of Nuclear Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu 610072, Sichuan Province, China
Xu-Fan Zhang, College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, Sichuan Province, China
Jia-Lian Zheng, Department of Hepatology, Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032, Liaoning Province, China
Zi-Yi Zhao, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu 610072, Sichuan Province, China
Zi-Yi Zhao, Traditional Chinese Medicine Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Chengdu 610072, Sichuan Province, China
Co-first authors: Yun Yuan and Xu-Fan Zhang.
Co-corresponding authors: Jia-Lian Zheng and Qiong-Ying Hu.
Author contributions: Yuan Y, Zhang XF, Li YC, Chen HQ, Wen T, Zheng JL, Zhao ZY, and Hu QY contributed to the investigation of this manuscript; Yuan Y, Li YC, Chen HQ, and Wen T were involved in the data curation; Yuan Y, Li YC, Zheng JL, and Hu QY participated in the methodology of this study; Yuan Y, Zhang XF, Zheng JL, Zhao ZY, and Hu QY took part in the conceptualization of this article; Yuan Y and Zhang XF wrote the original draft; Zhang XF, Zheng JL, and Hu QY contributed to the project administration of this manuscript; Zheng JL, Zhao ZY, and Hu QY were major in the funding acquisition and supervision of this program.
Supported by National Natural Science Foundation of China, No. 82074298; Chengdu Science and Technology Bureau Project, No. 2021-YF05-01726-SN; and “Xinglin Scholars” Research Promotion Program of Chengdu University of Traditional Chinese Medicine, No. QJRC2022007.
Institutional review board statement: The study was reviewed and approved by Related Ethical Regulations of Chengdu University of Traditional Chinese Medicine.
Institutional animal care and use committee statement: All the animal experiments were approved by the ethics committee of the Institutional Animal Care and Use Committee of Institute of Chengdu University of Traditional Chinese Medicine.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: All data during the study are available from the corresponding author by request at qiongyinghu@163.com.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Qiong-Ying Hu, PhD, Associate Professor, Department of Laboratory Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, No. 39 Shierqiao Road, Jinniu District, Chengdu 610072, Sichuan Province, China. qiongyinghu@163.com
Received: October 30, 2023
Peer-review started: October 30, 2023
First decision: December 5, 2023
Revised: December 19, 2023
Accepted: January 16, 2024
Article in press: January 16, 2024
Published online: February 26, 2024
Processing time: 118 Days and 15.4 Hours
Abstract
BACKGROUND

Colorectal cancer stem cells (CCSCs) are heterogeneous cells that can self-renew and undergo multidirectional differentiation in colorectal cancer (CRC) patients. CCSCs are generally accepted to be important sources of CRC and are responsible for the progression, metastasis, and therapeutic resistance of CRC. Therefore, targeting this specific subpopulation has been recognized as a promising strategy for overcoming CRC.

AIM

To investigate the effect of VX-509 on CCSCs and elucidate the underlying mechanism.

METHODS

CCSCs were enriched from CRC cell lines by in conditioned serum-free medium. Western blot, Aldefluor, transwell and tumorigenesis assays were performed to verify the phenotypic characteristics of the CCSCs. The anticancer efficacy of VX-509 was assessed in HCT116 CCSCs and HT29 CCSCs by performing cell viability analysis, colony formation, sphere formation, flow cytometry, and western blotting assessments in vitro and tumor growth, immunohistochemistry and immunofluorescence assessments in vivo.

RESULTS

Compared with parental cells, sphere cells derived from HCT116 and HT29 cells presented increased expression of stem cell transcription factors and stem cell markers and were more potent at promoting migration and tumorigenesis, demonstrating that the CRC sphere cells displayed CSC features. VX-509 inhibited the tumor malignant biological behavior of CRC-stem-like cells, as indicated by their proliferation, migration and clonality in vitro, and suppressed the tumor of CCSC-derived xenograft tumors in vivo. Besides, VX-509 suppressed the CSC characteristics of CRC-stem-like cells and inhibited the progression of epithelial-mesenchymal transition (EMT) signaling in vitro. Nodal was identified as the regulatory factor of VX-509 on CRC stem-like cells through analyses of differentially expressed genes and CSC-related database information. VX-509 markedly downregulated the expression of Nodal and its downstream phosphorylated Smad2/3 to inhibit EMT progression. Moreover, VX-509 reversed the dedifferentiation of CCSCs and inhibited the progression of EMT induced by Nodal overexpression.

CONCLUSION

VX-509 prevents the EMT process in CCSCs by inhibiting the transcription and protein expression of Nodal, and inhibits the dedifferentiated self-renewal of CCSCs.

Keywords: Colorectal cancer stem cells; Stemness; VX-509; Epithelial-mesenchymal transition; Nodal

Core Tip: Colorectal cancer (CRC) is a malignant cancer of the digestive tract with high recurrence and metastasis. CRC stem cells (CCSCs) exerted self-renewing and repopulating capacity, which are hard to eliminate and cause recurrence and metastasis of CRC. VX-509 has been reported as a rheumatoid arthritis regular treatment. However, the role of VX-509 in CCSCs is not well studied. In this study, VX-509 inhibited the malignant biological behaviors and stemness of CCSCs. Further experiments suggested that VX-509 downregulated the expression of Nodal to suppress the epithelial-mesenchymal transition and inhibit the stemness of CCSCs, which provided a potent therapy measure for CRC.