Research Paper Volume 12, Issue 17 pp 17662—17680
Shikonin overcomes drug resistance and induces necroptosis by regulating the miR-92a-1-5p/MLKL axis in chronic myeloid leukemia
- 1 Department of Hematology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China
- 2 Department of Hematology, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, China
- 3 Department of Hematology, Hangzhou First People’s Hospital, Hangzhou 310006, China
- 4 Department of Hematology, The Fourth Affiliated Hospital, College of Medicine, Zhejiang University, Yiwu 322000, China
Received: March 24, 2020 Accepted: July 7, 2020 Published: September 14, 2020
https://doi.org/10.18632/aging.103844How to Cite
Abstract
Development of resistance to tyrosine kinase inhibitors (TKIs) targeting the BCR/ABL fusion protein represents a major challenge in the treatment of chronic myeloid leukemia (CML). Since apoptosis resistance is the fundamental mechanism impeding TKIs’ therapeutic effects, alternative approaches that induce nonapoptotic cell death are being pursued to treat TKI-resistant CML. Induction of necroptosis, a distinct, caspase-independent form of programmed cell death, may be a valuable strategy in this respect. The present study shows that shikonin, an herbal compound used in traditional Chinese medicine, overcomes TKI resistance in BCR/ABL-positive CML cells by inducing necroptosis via activation of RIPK1/RIPK3/MLKL signaling. This effect occurs both in vitro and in vivo and involves downregulation of miR-92a-1-5p, a poor-prognosis marker frequently overexpressed in leukemia patients. Based on gene expression experiments, we conclude that miR-92a-1-5p promotes CML progression by inhibiting MLKL expression. Accordingly, we show that antagomiR-mediated in vivo inhibition of miR-92a-1-5p reduces the growth of CML tumors in mice through necroptosis induction. Our research suggests that therapies that relieve MLKL suppression by targeting miR-92a-1-5p may represent a useful strategy to treat TKI-refractory CML.
Abbreviations
CML: Chronic myelogenous leukemia; Ph: Philadelphia chromosome; BCR/ABL: Breakpoint Cluster Region-Abelson; TKI: Tyrosine kinase inhibitor; TNFα: Tumor necrosis factor α; TNFR: TNF receptor; TRAIL: TNF-related apoptosis-inducing ligand; FasL: Fas ligand; RIPK1/3: Receptor-interacting protein kinase-1, -3; MLKL: Mixed lineage kinase-domain like protein; ROS: Reactive oxygen species; miRNAs: microRNAs; NGS: Next-generation sequencing; Necrostatin-1: Nec-1; DMSO: Dimethyl sulfoxide; PI: Propidium iodide; MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; BCA: Bicinchoninic acid; SDS: Sodium dodecyl sulfate; HHT: Homoharringtonine; MMP: Mitochondrial membrane potential; MFI: Mean fluorescence intensity; NC: Negative control; IC50: half maximal inhibitory concentration; ISH: In situ hybridization; IHC: Immunohistochemistry; NOD/SCID mice: Nonobese diabetic/severe combined immunodeficiency mice; CTRL: control; SHK: shikonin; zVAD: zVAD-fmk; LV: lentivirus; CML-CP: CML-chronic phase; CML-BC: CML-blast crisis; IOD: integrated optical density.