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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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Chemical structure and antiproliferative activity against CCRF-CEM cells of acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a).(A) Chemical structure of 8a. (B) Viability of CCRF-CEM cells after 24 h of exposure to 8a. The viability of the control cells, which were exposed to DMSO only, was set as 100%. n = 6, *p<0.05 compared with vehicle control. (C) Flow cytometric analysis of phosphatidylserine externalization (annexin V-binding) and cell membrane integrity (PI staining). CCRF-CEM cells were treated with 8a at 0.5 µM for 24 h.
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pone-0063572-g001: Chemical structure and antiproliferative activity against CCRF-CEM cells of acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a).(A) Chemical structure of 8a. (B) Viability of CCRF-CEM cells after 24 h of exposure to 8a. The viability of the control cells, which were exposed to DMSO only, was set as 100%. n = 6, *p<0.05 compared with vehicle control. (C) Flow cytometric analysis of phosphatidylserine externalization (annexin V-binding) and cell membrane integrity (PI staining). CCRF-CEM cells were treated with 8a at 0.5 µM for 24 h.

Mentions: In our previous studies, a series of acridone derivatives have been synthesized, and among them, the derivative 8a, 2-aminoacetamido-10-(3,5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (Figure 1A), has shown potent antitumor activity. However, the underlying mechanism of the anti-proliferative activity of 8a is unclear [32]. This current study presented a global analysis of metabolic changes in CCRF-CEM leukemia cells treated with 9(10H)-acridone (A), 10-(3,5-dimethoxy)benzyl-9(10H)-acridone (I), and 8a (the optimized structure based on the former two compounds). Distinct metabolites were identified and the involved biological events were verified by conventional molecular biology.


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)

Chemical structure and antiproliferative activity against CCRF-CEM cells of acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a).(A) Chemical structure of 8a. (B) Viability of CCRF-CEM cells after 24 h of exposure to 8a. The viability of the control cells, which were exposed to DMSO only, was set as 100%. n = 6, *p<0.05 compared with vehicle control. (C) Flow cytometric analysis of phosphatidylserine externalization (annexin V-binding) and cell membrane integrity (PI staining). CCRF-CEM cells were treated with 8a at 0.5 µM for 24 h.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0063572-g001: Chemical structure and antiproliferative activity against CCRF-CEM cells of acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a).(A) Chemical structure of 8a. (B) Viability of CCRF-CEM cells after 24 h of exposure to 8a. The viability of the control cells, which were exposed to DMSO only, was set as 100%. n = 6, *p<0.05 compared with vehicle control. (C) Flow cytometric analysis of phosphatidylserine externalization (annexin V-binding) and cell membrane integrity (PI staining). CCRF-CEM cells were treated with 8a at 0.5 µM for 24 h.
Mentions: In our previous studies, a series of acridone derivatives have been synthesized, and among them, the derivative 8a, 2-aminoacetamido-10-(3,5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (Figure 1A), has shown potent antitumor activity. However, the underlying mechanism of the anti-proliferative activity of 8a is unclear [32]. This current study presented a global analysis of metabolic changes in CCRF-CEM leukemia cells treated with 9(10H)-acridone (A), 10-(3,5-dimethoxy)benzyl-9(10H)-acridone (I), and 8a (the optimized structure based on the former two compounds). Distinct metabolites were identified and the involved biological events were verified by conventional molecular biology.

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