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

Wang Y, Gao D, Chen Z, Li S, Gao C, Cao D, Liu F, Liu H, Jiang Y - PLoS ONE (2013)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Tsinghua University, Beijing, China.

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.

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PCA scores plot resulting from the UPLC/MS spectra of CCRF-CEM cells with corresponding OPLS-DA for tested groups.(A) PCA scores plot from control and 8a-treated group. (B) OPLS-DA scores plot from control and 8a-treated group. (C) Potential biomarkers in the S-plot between control and 8a-treated group.
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pone-0063572-g002: PCA scores plot resulting from the UPLC/MS spectra of CCRF-CEM cells with corresponding OPLS-DA for tested groups.(A) PCA scores plot from control and 8a-treated group. (B) OPLS-DA scores plot from control and 8a-treated group. (C) Potential biomarkers in the S-plot between control and 8a-treated group.

Mentions: To investigate the metabolic fingerprints in cancer cells, metabolites in CCRF-CEM cells from control and treated groups were profiled by UPLC/Q-TOF MS in both positive and negative modes, though positive ion mode gave more information-rich data than negative. Representative base peak intensity (BPI) chromatograms were shown in Figure S1. Raw data from UPLC/Q-TOF MS were analyzed by the Marker-Lynx software. The acquired QC data for UPLC/Q-TOF MS metabolic profiling were used to investigate the analytical variability in the whole run, which was critical for evaluating the variations in the analytical results and thus the reliability of the metabolite profiling data [37]. As shown in Figure 2A, the QC samples were tightly clustered in PCA scores plot, indicating the non-targeted analysis displayed the column stability in the whole run. Moreover, it was noteworthy that the control and 8a-treated groups were obviously separated along the first principal component, while A and I treated groups were not significantly different from the control group. It is indicated that the cellular metabolic phenotypes were significantly altered under the treatment of 8a. The results were in agreement with aforementioned anti-proliferative activity.


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

Wang Y, Gao D, Chen Z, Li S, Gao C, Cao D, Liu F, Liu H, Jiang Y - PLoS ONE (2013)

PCA scores plot resulting from the UPLC/MS spectra of CCRF-CEM cells with corresponding OPLS-DA for tested groups.(A) PCA scores plot from control and 8a-treated group. (B) OPLS-DA scores plot from control and 8a-treated group. (C) Potential biomarkers in the S-plot between control and 8a-treated group.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3646819&req=5

pone-0063572-g002: PCA scores plot resulting from the UPLC/MS spectra of CCRF-CEM cells with corresponding OPLS-DA for tested groups.(A) PCA scores plot from control and 8a-treated group. (B) OPLS-DA scores plot from control and 8a-treated group. (C) Potential biomarkers in the S-plot between control and 8a-treated group.
Mentions: To investigate the metabolic fingerprints in cancer cells, metabolites in CCRF-CEM cells from control and treated groups were profiled by UPLC/Q-TOF MS in both positive and negative modes, though positive ion mode gave more information-rich data than negative. Representative base peak intensity (BPI) chromatograms were shown in Figure S1. Raw data from UPLC/Q-TOF MS were analyzed by the Marker-Lynx software. The acquired QC data for UPLC/Q-TOF MS metabolic profiling were used to investigate the analytical variability in the whole run, which was critical for evaluating the variations in the analytical results and thus the reliability of the metabolite profiling data [37]. As shown in Figure 2A, the QC samples were tightly clustered in PCA scores plot, indicating the non-targeted analysis displayed the column stability in the whole run. Moreover, it was noteworthy that the control and 8a-treated groups were obviously separated along the first principal component, while A and I treated groups were not significantly different from the control group. It is indicated that the cellular metabolic phenotypes were significantly altered under the treatment of 8a. The results were in agreement with aforementioned anti-proliferative activity.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Tsinghua University, Beijing, China.

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.

Show MeSH
Related in: MedlinePlus