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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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Related in: MedlinePlus

The comparison of biomarker intensity between control and compounds-treated cells.These biomarkers involved in (A) Glutathione metabolism (B) Glycerophospholipid metabolism (n = 8), *p<0.05 and **p<0.01 compared with vehicle control.
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pone-0063572-g003: The comparison of biomarker intensity between control and compounds-treated cells.These biomarkers involved in (A) Glutathione metabolism (B) Glycerophospholipid metabolism (n = 8), *p<0.05 and **p<0.01 compared with vehicle control.

Mentions: On the basis of UPLC/Q-TOF MS metabolomics, identified variables from comparisons (8a, I or A-treated groups vs. control group, respectively) were summarized in Table S2, including RT, the mass obtained in UPLC-MS system, and the mass error when comparing with the database. In addition, the type of the identification (MS/MS fragmentation or confirmation with the analysis of standard), fragment ions of the metabolite, percentage of changes in different comparisons, and statistical significance were also presented in the table. It is shown that the metabolites in 8a-treated group were the most significantly changed. By comparing the 8a-treated group with the control, 23 metabolites were highlighted, which were derived from altered fatty acid, nucleoside, amino acid, glycerophospholipid, and glutathione metabolism. Further studies were focused on 11 metabolites which were involved in glutathione and glycerophospholipid metabolism. As shown in Figure 3, the metabolites in 8a, I and A-treated groups showed the same variation tendency, but extents of the changes in 8a-treated group were significantly larger than the other two groups. Therefore, the analysis of the changed metabolites, as well as the following biological studies, were focused on 8a-treated group.


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)

The comparison of biomarker intensity between control and compounds-treated cells.These biomarkers involved in (A) Glutathione metabolism (B) Glycerophospholipid metabolism (n = 8), *p<0.05 and **p<0.01 compared with vehicle control.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0063572-g003: The comparison of biomarker intensity between control and compounds-treated cells.These biomarkers involved in (A) Glutathione metabolism (B) Glycerophospholipid metabolism (n = 8), *p<0.05 and **p<0.01 compared with vehicle control.
Mentions: On the basis of UPLC/Q-TOF MS metabolomics, identified variables from comparisons (8a, I or A-treated groups vs. control group, respectively) were summarized in Table S2, including RT, the mass obtained in UPLC-MS system, and the mass error when comparing with the database. In addition, the type of the identification (MS/MS fragmentation or confirmation with the analysis of standard), fragment ions of the metabolite, percentage of changes in different comparisons, and statistical significance were also presented in the table. It is shown that the metabolites in 8a-treated group were the most significantly changed. By comparing the 8a-treated group with the control, 23 metabolites were highlighted, which were derived from altered fatty acid, nucleoside, amino acid, glycerophospholipid, and glutathione metabolism. Further studies were focused on 11 metabolites which were involved in glutathione and glycerophospholipid metabolism. As shown in Figure 3, the metabolites in 8a, I and A-treated groups showed the same variation tendency, but extents of the changes in 8a-treated group were significantly larger than the other two groups. Therefore, the analysis of the changed metabolites, as well as the following biological studies, were focused on 8a-treated group.

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