MLKL contributes to shikonin-induced glioma cell necroptosis via promotion of chromatinolysis.

Abstract

Chromatinolysis refers to enzymatic degradation of nuclear DNA and is regarded as one of the crucial events leading to cell death. Mixed-lineage kinase domain-like protein (MLKL) has been identified as a key executor of necroptosis, but it remains unclear whether MLKL contributes to necroptosis via regulation of chromatinolysis. In this study, we find that shikonin induces MLKL activation and chromatinolysis in glioma cells in vitro and in vivo, which are accompanied with nuclear translocation of AIF and γ-H2AX formation. In vitro studies reveal that inhibition of MLKL with its specific inhibitor NSA or knockdown of MLKL with siRNA abrogates shikonin-induced glioma cell necroptosis, as well as chromatinolysis. Mechanistically, activated MLKL targets mitochondria and triggers excessive generation of mitochondrial superoxide, which promotes AIF translocation into nucleus via causing mitochondrial depolarization and aggravates γ-H2AX formation via improving intracellular accumulation of ROS. Inhibition of nuclear level of AIF by knockdown of AIF with siRNA or mitigation of γ-H2AX formation by suppressing ROS with antioxidant NAC effectively prevents shikonin-induced chromatinolysis. Then, we found that RIP3 accounts for shikonin-induced activation of MLKL, and activated MLKL reversely up-regulates the protein level of CYLD and promotes the activation of RIP1 and RIP3. Taken together, our data suggest that MLKL contributes to shikonin-induced glioma cell necroptosis via promotion of chromatinolysis, and shikonin induces a positive feedback between MLKL and its upstream signals RIP1 and RIP3.

• Authors: Ye Ding, Chuan He, Shan Lu, Xuanzhong Wang, Chongcheng Wang, Lei Wang, Ji Zhang, Meihua Piao, Guangfan Chi, Yinan Luo, Ke Sai, Pengfei Ge
• Journal: Cancer letters (Cancer Lett) Vol. 467 Pg. 58-71 (12 28 2019) ISSN: 1872-7980 [Electronic] Ireland
• PMID: 31560934 (Publication Type: Journal Article)
• Copyright: Copyright © 2019 The Author(s). Published by Elsevier B.V. All rights reserved.

Chemical References

• AIFM1 protein, human
• Apoptosis Inducing Factor
• DNA, Neoplasm
• Naphthoquinones
• Reactive Oxygen Species
• shikonin
• MLKL protein, human
• Protein Kinases

Topics

• Animals
• Apoptosis Inducing Factor (metabolism)
• Cell Line, Tumor
• DNA Fragmentation
• DNA, Neoplasm (chemistry, drug effects)
• Gene Expression Regulation, Neoplastic (drug effects)
• Glioma (drug therapy, genetics, metabolism)
• Humans
• Mice
• Mitochondria (metabolism)
• Naphthoquinones (administration & dosage, pharmacology)
• Necroptosis
• Protein Kinases (metabolism)
• Rats
• Reactive Oxygen Species (metabolism)
• Xenograft Model Antitumor Assays