Shikonin as a WT1 Inhibitor Promotes Promyeloid Leukemia Cell Differentiation

• Published in: Molecules (Basel, Switzerland) Vol. 27; no. 23; p. 8264
• Main Authors: Guo, Zhenzhen, Sun, Luyao, Xia, Haojie, Tian, Shibin, Liu, Mengyue, Hou, Jiejie, Li, Jiahuan, Lin, Haihong, Du, Gangjun
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 26.11.2022; MDPI
• Subjects: Analysis; Animals; Antigens, CD34 - metabolism; Apoptosis; CD11b antigen; CD34; CD34 antigen; Cell cycle; Cell Differentiation; Cell growth; Cell proliferation; Cell self-renewal; Cell viability; Cytotoxicity; Deoxyribonucleic acid; Differentiation (biology); DNA; Drug dosages; Hydrogen peroxide; Hydrogen Peroxide - pharmacology; Leukemia; Leukemia - metabolism; Leukocytes (granulocytic); Localization; Medical prognosis; Membrane potential; Mice; Mitochondria; Molecular docking; Molecular Docking Simulation; Monocytes; Promyeloid leukemia; Protein binding; Protein interaction; Proteins; Shikonin; Tumors; Wilms’ tumor gene 1 (WT1); WT1 protein; WT1 Proteins - genetics; China; leukemia; cell differentiation; Wilms’ tumor gene 1 (WT1); CD34; shikonin
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules27238264

This study aims to observe the differentiating effect of shikonin on Wilms' tumor 1 (WT1)-positive HL-60 cells and investigate the fate of the differentiated leukemia cells. WT1 overexpression unaffected cell viability but promoted resistance to H O -induced DNA injury and cell apoptosis. The binding of shikonin to the WT1 protein was confirmed by molecular docking and drug affinity reaction target stability (DARTS). Shikonin at the non-cytotoxic concentration could decrease the WT1 protein and simultaneously reduced the CD34 protein and increased the CD11b protein in a dose-dependent manner in normal HL-60 cells but not in WT1-overexpressed HL-60 cells. Shikonin unaffected HL-60 cell viability in 48 h. However, it lasted for 10 days; could attenuate cell proliferation, mitochondrial membrane potential (MMP), and self-renewal; prevent the cell cycle; promote cell apoptosis. In a mouse leukemia model, shikonin could decrease the WT1 protein to prevent leukemia development in a dose-dependent manner. In this study, we also confirmed preliminarily the protein-protein interactions between WT1 and CD34 in molecular docking and CO-IP assay. Our results suggest that: 1. shikonin can down-regulate the WT1 protein level for leukemia differentiation therapy, and 2. the interaction between WT1 and CD34 proteins may be responsible for granulocyte/monocyte immaturity in HL-60 cells.