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Metabolomics Reveals Cysteine Metabolism Plays a Role in Celastrol-Induced Mitochondrial Apoptosis of Acute Promyelocytic Leukemia Cells

39 Pages Posted: 19 Feb 2019

See all articles by Minjian Chen

Minjian Chen

Nanjing Medical University - State Key Laboratory of Reproductive Medicine; Nanjing Medical University - Key Laboratory of Modern Toxicology

Jing Yang

Shanghai University of Traditional Chinese Medicine

Lei Li

Nanjing Medical University

Xiaomei Lu

Nanjing Medical University

Rongli Sun

Southeast University

Yubang Wang

Nanjing Medical University

Xiaoling Zhang

Nanjing Medical University - Department of Hygienic Analysis and Detection

More...

Abstract

Background: Recently, celastrol shows great potential for inducing apoptosis in the treatment of leukemia, especially acute promyelocytic leukemia. However, the mechanism is poorly understood. Metabolomics provides overall understanding of biological mechanisms by surveying small molecules, which contributes to the illustration of celastrol's action mode.  

Methods: We treated nude mice bearing xenografts of HL-60 cell in vivo and HL-60 cell as well as NB-4 cell in vitro with celastrol. UPLC coupled to an Orbitrap mass spectrometer was used for a hypothesis free metabolomics analysis of HL-60 in vivo, and was used for targeted L-cysteine analysis of HL-60 and NB-4 cells. Flow cytometry analysis was performed to detect the mitochondrial membrane potential, production of reactive oxygen species (ROS) and percentage of apoptosis. Western blot was conducted to detect the protein level of p53, Bax, cleaved caspase 9 and cleaved caspase 3.  

Findings: Celastrol inhibited tumor growth, induced apoptosis, and up-regulated pro-apoptotic proteins in xenograft tumor mice model. Metabolomics profiles delineated cysteine metabolism was the key changed metabolism after celastrol treatment in HL-60 cells in vivo. Celastrol decreased L-cysteine in HL-60 cells. The supplementation of acetylcysteine reversed the ROS and apoptosis induced by celastrol, and reversed the dramatic decline of mitochondrial membrane potential and the up-regulation of pro-apoptotic proteins in HL-60 cells. Celastrol decreased L-cysteine, and acetylcysteine reversed ROS and apoptosis induced by celastrol in NB-4 cells.  

Interpretation: We firstly identified cysteine metabolism/ROS/p53/Bax/caspase 9/caspase 3 pathway in celastrol-triggered apoptosis of APL cells, providing a novel mechanism underlying which celastrol might be used for the treatment of acute promyelocytic leukemia.  

Funding: This study was supported by grants from the National Natural Science Foundation of China (81872650, 81503134, 81573182); the Key Natural Science Foundation of the Jiangsu Higher Education Institutions of China (18KJA320003); the Key Project of the Science and Technology Development Foundation of Nanjing Medical University (2014NJMUZD005); the Key Research & Development Plan of Jiangsu Province (BE2017628); the Southeast University & Nanjing Medical University Collaborative Research Project (2242018K3DN25); the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).  

Declaration of Interest: The authors declare that they have no competing interests.

Ethical Approval: All procedures performed in studies involving animals were in accordance with the ethical standards of Nanjing Medical 419 University Institutional Animal Care and Use Committee.

Keywords: celastrol, metabolomics, cysteine, apoptosis, mitochondria

Suggested Citation

Chen, Minjian and Yang, Jing and Li, Lei and Lu, Xiaomei and Sun, Rongli and Wang, Yubang and Zhang, Xiaoling, Metabolomics Reveals Cysteine Metabolism Plays a Role in Celastrol-Induced Mitochondrial Apoptosis of Acute Promyelocytic Leukemia Cells (February 15, 2019). Available at SSRN: https://ssrn.com/abstract=3335016 or http://dx.doi.org/10.2139/ssrn.3335016

Minjian Chen

Nanjing Medical University - State Key Laboratory of Reproductive Medicine ( email )

Nanjing
China

Nanjing Medical University - Key Laboratory of Modern Toxicology ( email )

Nanjing
China

Jing Yang

Shanghai University of Traditional Chinese Medicine

No. 1200, Cailun Road
Shanghai, 201203
China

Lei Li

Nanjing Medical University

300 Guangzhou Road
Nanjing, Jiangsu 210029
China

Xiaomei Lu

Nanjing Medical University

300 Guangzhou Road
Nanjing, Jiangsu 210029
China

Rongli Sun

Southeast University

Sipailou 2#
Nanjing, Jiangsu Province 210096
China

Yubang Wang

Nanjing Medical University

300 Guangzhou Road
Nanjing, Jiangsu 210029
China

Xiaoling Zhang (Contact Author)

Nanjing Medical University - Department of Hygienic Analysis and Detection ( email )

Nanjing, 211166
China