8-Hydroxydaidzein Downregulates JAK/STAT, MMP, Oxidative Phosphorylation, and PI3K/AKT Pathways in K562 Cells

Pei-Shan Wu; Chih-Yang Wang; Pin-Shern Chen; Jui-Hsiang Hung; Jui-Hung Yen; Ming-Jiuan Wu

doi:10.3390/biomedicines9121907

8-Hydroxydaidzein Downregulates JAK/STAT, MMP, Oxidative Phosphorylation, and PI3K/AKT Pathways in K562 Cells

Biomedicines. 2021 Dec 14;9(12):1907. doi: 10.3390/biomedicines9121907.

Authors

Pei-Shan Wu¹, Chih-Yang Wang^{2

3}, Pin-Shern Chen^{1

4}, Jui-Hsiang Hung⁴, Jui-Hung Yen^{5

6}, Ming-Jiuan Wu^{1

4}

Affiliations

¹ Department of Applied Life Science and Health, Chia Nan University of Pharmacy and Science, Tainan 717, Taiwan.
² Ph.D. Program for Cancer Molecular Biology and Drug Discovery, Taipei Medical University, Taipei 11031, Taiwan.
³ Graduate Institute of Cancer Biology and Drug Discovery, Taipei Medical University, Taipei 11031, Taiwan.
⁴ Department of Biotechnology, Chia Nan University of Pharmacy and Science, Tainan 717, Taiwan.
⁵ Department of Molecular Biology and Human Genetics, Tzu Chi University, Hualien 970, Taiwan.
⁶ Institute of Medical Sciences, Tzu Chi University, Hualien 970, Taiwan.

Abstract

A metabolite isolated from fermented soybean, 8-hydroxydaidzein (8-OHD, 7,8,4'-trihydroxyisoflavone, NSC-678112), is widely used in ethnopharmacological research due to its anti-proliferative and anti-inflammatory effects. We reported previously that 8-OHD provoked reactive oxygen species (ROS) overproduction, and induced autophagy, apoptosis, breakpoint cluster region-Abelson murine leukemia viral oncogene (BCR-ABL) degradation, and differentiation in K562 human chronic myeloid leukemia (CML) cells. However, how 8-OHD regulates metabolism, the extracellular matrix during invasion and metastasis, and survival signaling pathways in CML remains largely unexplored. High-throughput technologies have been widely used to discover the therapeutic targets and pathways of drugs. Bioinformatics analysis of 8-OHD-downregulated differentially expressed genes (DEGs) revealed that Janus kinase/signal transducer and activator of transcription (JAK/STAT), matrix metalloproteinases (MMPs), c-Myc, phosphoinositide 3-kinase (PI3K)/AKT, and oxidative phosphorylation (OXPHOS) metabolic pathways were significantly altered by 8-OHD treatment. Western blot analyses validated that 8-OHD significantly downregulated cytosolic JAK2 and the expression and phosphorylation of STAT3 dose- and time-dependently in K562 cells. Zymography and transwell assays also confirmed that K562-secreted MMP9 and invasion activities were dose-dependently inhibited by 8-OHD after 24 h of treatment. RT-qPCR analyses verified that 8-OHD repressed metastasis and OXPHOS-related genes. In combination with DisGeNET, it was found that 8-OHD's downregulation of PI3K/AKT is crucial for controlling CML development. A STRING protein-protein interaction analysis further revealed that AKT and MYC are hub proteins for cancer progression. Western blotting revealed that AKT phosphorylation and nuclear MYC expression were significantly inhibited by 8-OHD. Collectively, this systematic investigation revealed that 8-OHD exerts anti-CML effects by downregulating JAK/STAT, PI3K/AKT, MMP, and OXPHOS pathways, and MYC expression. These results could shed new light on the development of 8-OHD for CML therapy.

Keywords: 8-hydroxydaidzein; AKT; JAK/STAT; K562; MMP; OXPHOS.

Grants and funding

MOST 106-2320-B-041-006-MY3; MOST 110-2320-B-041-002-MY3; MOST 109-2320-B-038-009-MY2/Ministry of Science and Technology Taiwan