Open Access

ARTICLE

ICAT mediates the inhibition of stemness and tumorigenesis in acute myeloid leukemia cells induced by 1,25-(OH)₂D₃

YULIAN WANG¹, LIANLI ZHU¹, RONGHAO ZENG², YUNPING PU³, BAIJIAN CHEN³, YUWEI TAN³, MING HONG^2,*, WEIJIA WANG^1,2,*

1 Graduate School, Zhuhai Campus of Zunyi Medical University, Zhuhai, 519041, China
2 Department of Advanced Diagnostic and Clinical Medicine, Zhongshan People’s Hospital, Zhongshan, 528403, China
3 Graduate School, Guangdong Medical University, Zhanjiang, 524023, China

* Corresponding Authors: MING HONG. Email: ; WEIJIA WANG. Email:

Oncology Research 2025, 33(3), 695-708. https://doi.org/10.32604/or.2024.051746

Received 14 March 2024; Accepted 06 June 2024; Issue published 28 February 2025

Abstract

Background: The role of 1,25-dihydroxyvitamin D3 (1,25-(OH)₂D₃) in cancer prevention and treatment is an emerging topic of interest. However, its effects on the stemness of acute myeloid leukemia (AML) cells are poorly understood. Methods: The proliferation and differentiation of AML cells (HL60 and NB4) were investigated by the CCK-8 assay, immunocytochemical staining, and flow cytometry. The abilities of HL60 and NB4 cells to form spheres were examined by the cell sphere formation assay. In addition, the levels of stemness-associated markers (SOX2, Nanog, OCT4, and c-Myc) in HL60 and NB4 cells were measured by western blotting and quantitative real-time polymerase chain reaction. Moreover, we obtained β-catenin-interacting protein 1 (ICAT)-knockout and ICAT-overexpressing HL-60 cells using gene editing and lentiviral infection techniques and investigated the role of ICAT in modulating the stemness-inhibiting effects of 1,25-(OH)₂D₃ using the aforementioned experimental methods. Finally, we validated our findings in vivo using NOD/SCID mice. Results: 1,25-(OH)₂D₃ inhibited the proliferation and stemness of AML cells (HL60 and NB4) and induced their differentiation into monocytes. Additionally, the knockdown of ICAT in HL60 cells attenuated the inhibitory effects of 1,25-(OH)₂D₃ on proliferation and stemness and suppressed the expression of stemness markers. Conversely, overexpression of ICAT enhanced the aforementioned inhibitory effects of 1,25-(OH)₂D₃. Consistently, in NOD/SCID mice, 1,25-(OH)₂D₃ suppressed tumor formation by HL-60 cells, and the effects of ICAT knockdown or overexpression on 1,25-(OH)₂D₃ aligned with the in vitro findings. Conclusion: 1,25-(OH)₂D₃ inhibits AML cell stemness, possibly through modulation of the ICAT-mediated Wnt/β-catenin signaling pathway.

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