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Apoptotic Cell Death Induced by Resveratrol Is Partially Mediated by the Autophagy Pathway in Human Ovarian Cancer Cells.

Lang F, Qin Z, Li F, Zhang H, Fang Z, Hao E - PLoS ONE (2015)

Bottom Line: However, the mechanism of resveratrol-induced anti-cancer effect at the molecular level remains to be elucidated.The apoptotic cell death induced by resveratrol was attenuated by both pharmacological and genetic inhibition of autophagy.We also demonstrated that targeting ATG5 by siRNA also suppressed resveratrol-induced apoptotic cell death.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Jinan Central Hospital, Affiliated with Shandong University, Jinan, China.

ABSTRACT
Resveratrol (trans-3,4,5'-trihydroxystilbene) is an active compound in food, such as red grapes, peanuts, and berries. Resveratrol exhibits an anticancer effect on various human cancer cells. However, the mechanism of resveratrol-induced anti-cancer effect at the molecular level remains to be elucidated. In this study, the mechanism underlying the anti-cancer effect of resveratrol in human ovarian cancer cells (OVCAR-3 and Caov-3) was investigated using various molecular biology techniques, such as flow cytometry, western blotting, and RNA interference, with a major focus on the potential role of autophagy in resveratrol-induced apoptotic cell death. We demonstrated that resveratrol induced reactive oxygen species (ROS) generation, which triggers autophagy and subsequent apoptotic cell death. Resveratrol induced ATG5 expression and promoted LC3 cleavage. The apoptotic cell death induced by resveratrol was attenuated by both pharmacological and genetic inhibition of autophagy. The autophagy inhibitor chloroquine, which functions at the late stage of autophagy, significantly reduced resveratrol-induced cell death and caspase 3 activity in human ovarian cancer cells. We also demonstrated that targeting ATG5 by siRNA also suppressed resveratrol-induced apoptotic cell death. Thus, we concluded that a common pathway between autophagy and apoptosis exists in resveratrol-induced cell death in OVCAR-3 human ovarian cancer cells.

No MeSH data available.


Related in: MedlinePlus

Pharmacological inhibition of autophagy protects ovarian cancer cells OVCAR-3 against resveratrol-induced cell death.A. Representative flow cytometry dot diagrams representing each OVCAR-3 cells. Cellular apoptosis was measured in the same method as in Fig 1. Chloroquine reduced the cell killing effect of resveratrol. B. Quantitative determination of total cell death, which was expressed as the percentage of total cells. * P<0.05 compared with vehicle control; # P<0.05 compared with resveratrol-treated samples. n = 4/group. C. Caspase 3 and PARP activities were assayed and plotted as fold change. D. Representative western blotting analyses of OVCAR-3 total cell lysates with LC3, and p62 antibodies.
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pone.0129196.g006: Pharmacological inhibition of autophagy protects ovarian cancer cells OVCAR-3 against resveratrol-induced cell death.A. Representative flow cytometry dot diagrams representing each OVCAR-3 cells. Cellular apoptosis was measured in the same method as in Fig 1. Chloroquine reduced the cell killing effect of resveratrol. B. Quantitative determination of total cell death, which was expressed as the percentage of total cells. * P<0.05 compared with vehicle control; # P<0.05 compared with resveratrol-treated samples. n = 4/group. C. Caspase 3 and PARP activities were assayed and plotted as fold change. D. Representative western blotting analyses of OVCAR-3 total cell lysates with LC3, and p62 antibodies.

Mentions: To test whether autophagy is a link to apoptotic cell death, we treated OVCAR-3 cells with chloroquine (CQ), a pharmacological inhibitor of autophagy, prior to resveratrol treatment. Apoptosis and cell death were determined using Sytox Green and AnnexinV–Allophycocyanin. As shown in Fig 6A, cell death by resveratrol at 30 μM was significantly attenuated by pretreatment with 2 μM of CQ. Resveratrol significantly reduced the total cellular death by autophagy inhibition. also analyzed resveratrol induced apoptosis markers caspase 3 activity and PARP activity and both were attenuated by CQ treatment (Fig 6C). In addition, we observed that resveratrol-induced autophagosome markers LC3II and p62 were also attenuated by CQ treatment (Fig 6D).


Apoptotic Cell Death Induced by Resveratrol Is Partially Mediated by the Autophagy Pathway in Human Ovarian Cancer Cells.

Lang F, Qin Z, Li F, Zhang H, Fang Z, Hao E - PLoS ONE (2015)

Pharmacological inhibition of autophagy protects ovarian cancer cells OVCAR-3 against resveratrol-induced cell death.A. Representative flow cytometry dot diagrams representing each OVCAR-3 cells. Cellular apoptosis was measured in the same method as in Fig 1. Chloroquine reduced the cell killing effect of resveratrol. B. Quantitative determination of total cell death, which was expressed as the percentage of total cells. * P<0.05 compared with vehicle control; # P<0.05 compared with resveratrol-treated samples. n = 4/group. C. Caspase 3 and PARP activities were assayed and plotted as fold change. D. Representative western blotting analyses of OVCAR-3 total cell lysates with LC3, and p62 antibodies.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4466135&req=5

pone.0129196.g006: Pharmacological inhibition of autophagy protects ovarian cancer cells OVCAR-3 against resveratrol-induced cell death.A. Representative flow cytometry dot diagrams representing each OVCAR-3 cells. Cellular apoptosis was measured in the same method as in Fig 1. Chloroquine reduced the cell killing effect of resveratrol. B. Quantitative determination of total cell death, which was expressed as the percentage of total cells. * P<0.05 compared with vehicle control; # P<0.05 compared with resveratrol-treated samples. n = 4/group. C. Caspase 3 and PARP activities were assayed and plotted as fold change. D. Representative western blotting analyses of OVCAR-3 total cell lysates with LC3, and p62 antibodies.
Mentions: To test whether autophagy is a link to apoptotic cell death, we treated OVCAR-3 cells with chloroquine (CQ), a pharmacological inhibitor of autophagy, prior to resveratrol treatment. Apoptosis and cell death were determined using Sytox Green and AnnexinV–Allophycocyanin. As shown in Fig 6A, cell death by resveratrol at 30 μM was significantly attenuated by pretreatment with 2 μM of CQ. Resveratrol significantly reduced the total cellular death by autophagy inhibition. also analyzed resveratrol induced apoptosis markers caspase 3 activity and PARP activity and both were attenuated by CQ treatment (Fig 6C). In addition, we observed that resveratrol-induced autophagosome markers LC3II and p62 were also attenuated by CQ treatment (Fig 6D).

Bottom Line: However, the mechanism of resveratrol-induced anti-cancer effect at the molecular level remains to be elucidated.The apoptotic cell death induced by resveratrol was attenuated by both pharmacological and genetic inhibition of autophagy.We also demonstrated that targeting ATG5 by siRNA also suppressed resveratrol-induced apoptotic cell death.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Jinan Central Hospital, Affiliated with Shandong University, Jinan, China.

ABSTRACT
Resveratrol (trans-3,4,5'-trihydroxystilbene) is an active compound in food, such as red grapes, peanuts, and berries. Resveratrol exhibits an anticancer effect on various human cancer cells. However, the mechanism of resveratrol-induced anti-cancer effect at the molecular level remains to be elucidated. In this study, the mechanism underlying the anti-cancer effect of resveratrol in human ovarian cancer cells (OVCAR-3 and Caov-3) was investigated using various molecular biology techniques, such as flow cytometry, western blotting, and RNA interference, with a major focus on the potential role of autophagy in resveratrol-induced apoptotic cell death. We demonstrated that resveratrol induced reactive oxygen species (ROS) generation, which triggers autophagy and subsequent apoptotic cell death. Resveratrol induced ATG5 expression and promoted LC3 cleavage. The apoptotic cell death induced by resveratrol was attenuated by both pharmacological and genetic inhibition of autophagy. The autophagy inhibitor chloroquine, which functions at the late stage of autophagy, significantly reduced resveratrol-induced cell death and caspase 3 activity in human ovarian cancer cells. We also demonstrated that targeting ATG5 by siRNA also suppressed resveratrol-induced apoptotic cell death. Thus, we concluded that a common pathway between autophagy and apoptosis exists in resveratrol-induced cell death in OVCAR-3 human ovarian cancer cells.

No MeSH data available.


Related in: MedlinePlus