Acridone derivative 8a induces oxidative stress-mediated apoptosis in CCRF-CEM leukemia cells: application of metabolomics in mechanistic studies of antitumor agents

PLoS One. 2013 May 7;8(5):e63572. doi: 10.1371/journal.pone.0063572. Print 2013.

Abstract

A new acridone derivative, 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (named 8a) synthesized in our lab shows potent antitumor activity, but the mechanism of action remains unclear. Herein, we report the use of an UPLC/Q-TOF MS metabolomic approach to study the effects of three compounds with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy)benzyl-9(10H)-acridone (I), and 8a, on CCRF-CEM leukemia cells and to shed new light on the probable antitumor mechanism of 8a. Acquired data were processed by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) to identify potential biomarkers. Comparing 8a-treated CCRF-CEM leukemia cells with vehicle control (DMSO), 23 distinct metabolites involved in five metabolic pathways were identified. Metabolites from glutathione (GSH) and glycerophospholipid metabolism were investigated in detail, and results showed that GSH level and the reduced/oxidized glutathione (GSH/GSSG) ratio were significantly decreased in 8a-treated cells, while L-cysteinyl-glycine (L-Cys-Gly) and glutamate were greatly increased. In glycerophospholipid metabolism, cell membrane components phosphatidylcholines (PCs) were decreased in 8a-treated cells, while the oxidative products lysophosphatidylcholines (LPCs) were significantly increased. We further found that in 8a-treated cells, the reactive oxygen species (ROS) and lipid peroxidation product malondialdehyde (MDA) were notably increased, accompanied with decrease of mitochondrial transmembrane potential, release of cytochrome C and activation of caspase-3. Taken together our results suggest that the acridone derivative 8a induces oxidative stress-mediated apoptosis in CCRF-CEM leukemia cells. The UPLC/Q-TOF MS based metabolomic approach provides novel insights into the mechanistic studies of antitumor drugs from a point distinct from traditional biological investigations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acridines / chemistry
  • Acridines / pharmacology*
  • Acridones / chemistry
  • Acridones / pharmacology*
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects*
  • Biomarkers / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Chromatography, Liquid
  • Discriminant Analysis
  • Humans
  • Least-Squares Analysis
  • Leukemia / metabolism*
  • Leukemia / pathology*
  • Malondialdehyde / metabolism
  • Mass Spectrometry
  • Metabolomics*
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Models, Biological
  • Oxidative Stress / drug effects*
  • Principal Component Analysis
  • Reactive Oxygen Species / metabolism

Substances

  • 2-aminoacetamido-10-(3, 5-dimethoxy)benzyl-9(10H)-acridone
  • Acridines
  • Acridones
  • Antineoplastic Agents
  • Biomarkers
  • Reactive Oxygen Species
  • Malondialdehyde
  • acridone

Grant support

The research work was supported in part by the Chinese National Natural Science Foundation of China (Nos. 21172129), a finance supported by Chinese National Natural Science Foundation of China (20902053), China Postdoctoral Science Foundation (number 2012M510407), China Ministry of Science and Technology under Contract (number 2012ZX09506001-010), and International S&T Cooperation Program of China (number 2011DFA30620). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.