WYE-354 restores Adriamycin sensitivity in multidrug-resistant acute myeloid leukemia cell lines

Abstract

Multidrug resistance (MDR) is a major reason for the failure of acute myeloid leukemia (AML) therapy. Agents that reverse MDR and sensitize AML cells to chemotherapy are of great clinical significance. The present study developed Adriamycin (Adr)-resistant cell lines, namely K562/Adr200 and K562/Adr500, which exhibited MDR. The upregulation of ATP-binding cassette subfamily B member 1 (ABCB1) was confirmed as the mechanism of resistance by reverse transcription-quantitative polymerase chain reaction and western blot analyses. Subsequently, the role of the mammalian target of rapamycin (mTOR) kinase inhibitor, WYE-354, in sensitizing the K562/Adr200 and K562/Adr500 cell lines to Adr was evaluated. At sub-cytotoxic concentrations, WYE-354 increased Adr cytotoxicity in the K562/Adr200 and K562/Adr500 cells. WYE-354 restored Adr sensitivity in the resistant cells by inhibiting ABCB1-mediated substrate efflux, thereby leading to an accumulation of Adr, an increase in Adr-mediated G2/M cell cycle arrest and the induction of apoptosis. Furthermore, WYE-354 stimulated the ATPase activity of ABCB1, which was consistent with in silico predictions using a human ABCB1 mouse homology model, indicating that WYE-354 is a potent substrate of ABCB1. WYE-354 did not regulate the expression of ABCB1 at the concentrations used in the present study. These findings indicate that WYE-354 may be a competitive inhibitor of ABCB1-mediated efflux and a potential candidate in combination with standard chemotherapy for overcoming MDR. Further clinical investigations are warranted to validate this combination in vivo .

Overexpression of ABCB1 Transporter Confers Resistance to mTOR Inhibitor WYE-354 in Cancer Cells

“…However, in order to achieve the reversal effect, the dosage of verapamil often exceeds the safe dose in vivo, which hinders the application of such drugs in a clinical setting [40]. WYE-354 is an effective and specific mTOR inhibitor, which has recently reported to reverse ABCB1-mediated MDR [29,41]. To our knowledge, this study is the first to provide strong evidence that WYE-354 is a substrate of ABCB1, which indicates that warning and precaution should be given, especially when WYE-354 is used as an anticancer agent.…”

“…Moreover, the residues included in the interactions of human ABCB1 with WYE-354 and doxorubicin are different. Compared with the hABCB1 mouse homology model reported by Ibrahim et al [29], the human ABCB1 model we used for the molecular docking analysis is more suitable for our study. Since we used human cancer cells for the assays, the human ABCB1 model can better illustrate the interaction of WYE-354 and ABCB1 within human cancer cells.…”

“…The binding mode of WYE-354 with hABCB1 mouse homology model has been reported by Ibrahim et al [29]. In this study, we conducted the molecular docking analysis using human ABCB1 model (PDB 6QEX), which was published recently.…”

“…WYE-354 inhibits protein synthesis, cell growth, and division, and has an anticancer activity by inhibiting the phosphorylation of the related downstream proteins of mTORc2, and selectively inhibiting p-AKT (S473) via an mTORc2 target in different cancer types [28]. In addition, WYE-354 can reverse ABCB1-mediated MDR [29], but whether WYE-354 is a substrate of ABCB1 is not clear. Herein, in vitro experiments were conducted to evaluate whether WYE-354 is a substrate of the ABCB1 transporter.…”

In vitro Evaluation of the Anti-inflammatory Effects of Thymoquinone in Osteoarthritis and in silico Analysis of Inter-Related Pathways in Age-Related Degenerative Diseases

“…The fluorescence was measured at 590 nm using the SpectraMax ® i3x Multi-Mode microplate reader (Molecular Devices, LLC, San Jose, CA, United States). The values were plotted against drug concentrations and IC50 was determined ( Ibrahim et al, 2019 ).…”