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Pterostilbene exerts antitumor activity via the Notch1 signaling pathway in human lung adenocarcinoma cells.

Yang Y, Yan X, Duan W, Yan J, Yi W, Liang Z, Wang N, Li Y, Chen W, Yu S, Jin Z, Yi D - PLoS ONE (2013)

Bottom Line: PTE treatment resulted in a dose- and time-dependent decrease in the viability of A549 cells.DAPT (a gamma secretase inhibitor) and Notch1 siRNA prevented the induction of NICD and Hes1 activation by PTE treatment and sensitized the cells to PTE treatment.The down-regulation of Notch signaling also prevented the activation of pro-survival pathways (most notably the PI3K/Akt pathway) after PTE treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an City, China.

ABSTRACT
Although pterostilbene (PTE) has been shown to have potent antitumor activities against various cancer types, the molecular mechanisms of these activities remain unclear. In this study, we investigated the antitumor activity of PTE against human lung adenocarcinoma in vitro and in vivo and explored the role of the Notch1 signaling pathway in this process. PTE treatment resulted in a dose- and time-dependent decrease in the viability of A549 cells. Additionally, PTE exhibited strong antitumor activity, as evidenced not only by a reduced mitochondrial membrane potential (MMP) and a decreased intracellular glutathione content but also by increases in the apoptotic index and the level of reactive oxygen species (ROS). Furthermore, PTE treatment induced the activation of the Notch1 Intracellular Domain (NICD) protein and activated Hes1. DAPT (a gamma secretase inhibitor) and Notch1 siRNA prevented the induction of NICD and Hes1 activation by PTE treatment and sensitized the cells to PTE treatment. The down-regulation of Notch signaling also prevented the activation of pro-survival pathways (most notably the PI3K/Akt pathway) after PTE treatment. In summary, lung adenocarcinoma cells may enhance Notch1 activation as a protective mechanism in response to PTE treatment. Combining a gamma secretase inhibitor with PTE treatment may represent a novel approach for treating lung adenocarcinoma by inhibiting the survival pathways of cancer cells.

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The effects of PTE on ROS generation, the GSH level and the GSH/GSSG ratio in lung adenocarcinoma cells treated for 24 h.(A) To determine the ROS concentration, the cells were loaded with DCFH-DA, and the fluorescence intensity was measured using a microplate fluorescence reader. The fluorescence intensity in the control group was defined as 100%. (B) The intracellular GSH content was measured using a GSH kit. The differences in the GSH level between groups were expressed as a percentage of the control. (C) The GSH and GSSG levels were evaluated by comparison with the standards and were normalized to the protein content. The results are expressed as the GSH/GSSG ratio. The results are expressed as the mean ± SEM, n = 6. **P<0.01 compared with the control group, ##P<0.01 compared with the PTE 1.5 µM group, $$P<0.01 compared with the PTE 3 µM group. PTE, pterostilbene; ROS, reactive oxygen species; GSH, glutathione; GSSG, glutathione disulfide.
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pone-0062652-g003: The effects of PTE on ROS generation, the GSH level and the GSH/GSSG ratio in lung adenocarcinoma cells treated for 24 h.(A) To determine the ROS concentration, the cells were loaded with DCFH-DA, and the fluorescence intensity was measured using a microplate fluorescence reader. The fluorescence intensity in the control group was defined as 100%. (B) The intracellular GSH content was measured using a GSH kit. The differences in the GSH level between groups were expressed as a percentage of the control. (C) The GSH and GSSG levels were evaluated by comparison with the standards and were normalized to the protein content. The results are expressed as the GSH/GSSG ratio. The results are expressed as the mean ± SEM, n = 6. **P<0.01 compared with the control group, ##P<0.01 compared with the PTE 1.5 µM group, $$P<0.01 compared with the PTE 3 µM group. PTE, pterostilbene; ROS, reactive oxygen species; GSH, glutathione; GSSG, glutathione disulfide.

Mentions: To measure the capacity of PTE to cause intracellular oxidation, the specific oxidation-sensitive fluorescent dye DCFH-DA was used. The fluorescent intensity of this dye is increased following the generation of intracellular reactive metabolites. The treatment of A549 cells with PTE (1.5, 3 or 6 µM) for 24 h induced a dose-dependent increase in ROS generation (P<0.01, compared with the control group, Figure 3A).


Pterostilbene exerts antitumor activity via the Notch1 signaling pathway in human lung adenocarcinoma cells.

Yang Y, Yan X, Duan W, Yan J, Yi W, Liang Z, Wang N, Li Y, Chen W, Yu S, Jin Z, Yi D - PLoS ONE (2013)

The effects of PTE on ROS generation, the GSH level and the GSH/GSSG ratio in lung adenocarcinoma cells treated for 24 h.(A) To determine the ROS concentration, the cells were loaded with DCFH-DA, and the fluorescence intensity was measured using a microplate fluorescence reader. The fluorescence intensity in the control group was defined as 100%. (B) The intracellular GSH content was measured using a GSH kit. The differences in the GSH level between groups were expressed as a percentage of the control. (C) The GSH and GSSG levels were evaluated by comparison with the standards and were normalized to the protein content. The results are expressed as the GSH/GSSG ratio. The results are expressed as the mean ± SEM, n = 6. **P<0.01 compared with the control group, ##P<0.01 compared with the PTE 1.5 µM group, $$P<0.01 compared with the PTE 3 µM group. PTE, pterostilbene; ROS, reactive oxygen species; GSH, glutathione; GSSG, glutathione disulfide.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3643961&req=5

pone-0062652-g003: The effects of PTE on ROS generation, the GSH level and the GSH/GSSG ratio in lung adenocarcinoma cells treated for 24 h.(A) To determine the ROS concentration, the cells were loaded with DCFH-DA, and the fluorescence intensity was measured using a microplate fluorescence reader. The fluorescence intensity in the control group was defined as 100%. (B) The intracellular GSH content was measured using a GSH kit. The differences in the GSH level between groups were expressed as a percentage of the control. (C) The GSH and GSSG levels were evaluated by comparison with the standards and were normalized to the protein content. The results are expressed as the GSH/GSSG ratio. The results are expressed as the mean ± SEM, n = 6. **P<0.01 compared with the control group, ##P<0.01 compared with the PTE 1.5 µM group, $$P<0.01 compared with the PTE 3 µM group. PTE, pterostilbene; ROS, reactive oxygen species; GSH, glutathione; GSSG, glutathione disulfide.
Mentions: To measure the capacity of PTE to cause intracellular oxidation, the specific oxidation-sensitive fluorescent dye DCFH-DA was used. The fluorescent intensity of this dye is increased following the generation of intracellular reactive metabolites. The treatment of A549 cells with PTE (1.5, 3 or 6 µM) for 24 h induced a dose-dependent increase in ROS generation (P<0.01, compared with the control group, Figure 3A).

Bottom Line: PTE treatment resulted in a dose- and time-dependent decrease in the viability of A549 cells.DAPT (a gamma secretase inhibitor) and Notch1 siRNA prevented the induction of NICD and Hes1 activation by PTE treatment and sensitized the cells to PTE treatment.The down-regulation of Notch signaling also prevented the activation of pro-survival pathways (most notably the PI3K/Akt pathway) after PTE treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an City, China.

ABSTRACT
Although pterostilbene (PTE) has been shown to have potent antitumor activities against various cancer types, the molecular mechanisms of these activities remain unclear. In this study, we investigated the antitumor activity of PTE against human lung adenocarcinoma in vitro and in vivo and explored the role of the Notch1 signaling pathway in this process. PTE treatment resulted in a dose- and time-dependent decrease in the viability of A549 cells. Additionally, PTE exhibited strong antitumor activity, as evidenced not only by a reduced mitochondrial membrane potential (MMP) and a decreased intracellular glutathione content but also by increases in the apoptotic index and the level of reactive oxygen species (ROS). Furthermore, PTE treatment induced the activation of the Notch1 Intracellular Domain (NICD) protein and activated Hes1. DAPT (a gamma secretase inhibitor) and Notch1 siRNA prevented the induction of NICD and Hes1 activation by PTE treatment and sensitized the cells to PTE treatment. The down-regulation of Notch signaling also prevented the activation of pro-survival pathways (most notably the PI3K/Akt pathway) after PTE treatment. In summary, lung adenocarcinoma cells may enhance Notch1 activation as a protective mechanism in response to PTE treatment. Combining a gamma secretase inhibitor with PTE treatment may represent a novel approach for treating lung adenocarcinoma by inhibiting the survival pathways of cancer cells.

Show MeSH
Related in: MedlinePlus