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.
Highlights
The bioinformatics data revealed that 8-OHD downregulated JAK/STAT, phosphoinositide 3-kinase (PI3K)/AKT, matrix metalloproteinases (MMPs), and oxidative phosphorylation (OXPHOS) pathways in K562 cells
Validated data from Western blotting revealed that cytosolic JAK2 expression and phosphorylation of STAT3 decreased in dose-dependent manners in 8-OHD-treated K562 cells
The repression of MMP14, MMP15, VEGFA, Son of Sevenless 1 (SOS1), and MMP11 by 8-OHD may indicate that it possesses anti-metastatic activity
Summary
Leukemia is a hematologic disorder, which can be classified into four main types, including acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), and chronic myeloid leukemia (CML), according to the speed. A decrease in phosphorylated STAT3 and an undetectable BCR-AB1 transcript were observed after treatment with ruxolitinib (INCB18424), a JAK inhibitor, in combination with the TKI, nilotinib, in CML patients in a phase I trial (NCT01702064) [10]. It has become clear that TKI-resistant AML and CML, and leukemic stem cells (LSCs) are accompanied by increased OXPHOS, regardless of the presence or absence of genetic mutations; interest has grown in developing clinically applicable OXPHOS inhibitors [16] It is well-known that mitochondria play key roles in activating apoptosis in mammalian cells and B-cell lymphoma protein 2 (Bcl-2) family members are the key regulators thereof [17]. We investigated the anti-CML effects of 8-OHD, focusing on cell-cycle regulation, cell apoptosis, autophagy, differentiation, and the altered level of BCR-ABL in K562 cells [39]. These results shed new light on potential therapeutic targets for CML treatment
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
