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Induction of apoptosis and autophagy via sirtuin1- and PI3K/Akt/mTOR-mediated pathways by plumbagin in human prostate cancer cells.

Zhou ZW, Li XX, He ZX, Pan ST, Yang Y, Zhang X, Chow K, Yang T, Qiu JX, Zhou Q, Tan J, Wang D, Zhou SF - Drug Des Devel Ther (2015)

Bottom Line: Our study has shown that PLB had potent pro-apoptotic and pro-autophagic effects on PC-3 and DU145 cells.In addition, PLB downregulated pre-B cell colony-enhancing factor/visfatin, and the inhibition of pre-B cell colony-enhancing factor/visfatin significantly enhanced basal and PLB-induced apoptosis and autophagy in both cell lines.Moreover, reduction of intracellular reactive oxygen species (ROS) level attenuated the apoptosis- and autophagy-inducing effects of PLB on both PC-3 and DU145 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA ; Guizhou Provincial Key Laboratory for Regenerative Medicine, Stem Cell and Tissue Engineering Research Center and Sino-US Joint Laboratory for Medical Sciences, Guizhou Medical University, Guiyang, Guizhou, People's Republic of China.

ABSTRACT
Plumbagin (PLB) has been shown to have anticancer activities in animal models, but the role of PLB in prostate cancer treatment is unclear. This study aimed to investigate the effects of PLB on apoptosis and autophagy and the underlying mechanisms in human prostate cancer cell lines PC-3 and DU145. Our study has shown that PLB had potent pro-apoptotic and pro-autophagic effects on PC-3 and DU145 cells. PLB induced mitochondria-mediated apoptosis and autophagy in concentration- and time-dependent manners in both PC-3 and DU145 cells. PLB induced inhibition of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) and p38 mitogen-activated protein kinase (MAPK) pathways and activation of 5'-AMP-dependent kinase (AMPK) as indicated by their altered phosphorylation, contributing to the pro-autophagic activity of PLB. Modulation of autophagy altered basal and PLB-induced apoptosis in both cell lines. Furthermore, PLB downregulated sirtuin 1 (Sirt1), and inhibition of Sirt1 enhanced autophagy, whereas the induction of Sirt1 abolished PLB-induced autophagy in PC-3 and DU145 cells. In addition, PLB downregulated pre-B cell colony-enhancing factor/visfatin, and the inhibition of pre-B cell colony-enhancing factor/visfatin significantly enhanced basal and PLB-induced apoptosis and autophagy in both cell lines. Moreover, reduction of intracellular reactive oxygen species (ROS) level attenuated the apoptosis- and autophagy-inducing effects of PLB on both PC-3 and DU145 cells. These findings indicate that PLB promotes apoptosis and autophagy in prostate cancer cells via Sirt1- and PI3K/Akt/mTOR-mediated pathways with contribution from AMPK-, p38 MAPK-, visfatin-, and ROS-associated pathways.

No MeSH data available.


Related in: MedlinePlus

ROS plays a role in PLB-induced apoptosis and autophagy in PC-3 and DU145 cells.Notes: (A) PLB stimulating the generation of ROS in PC-3 and DU145 cells. Cells were treated with PLB at 0.1 μM, 1 μM, or 5 μM for 24 hours. The bar graphs showing the intracellular level of ROS in PC-3 and DU145 cells. (B) Bar graphs showing the time course of PLB-induced generation of intracellular ROS in PC-3 and DU145 cells. (C) Effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (D) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (E) Representative flow cytometric dot plots showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells. (F) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells determined by flow cytometry. (G) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (H) Bar graphs showing the effects of CDDO-Me on basal and PBL-induced apoptosis in PC-3 and DU145 cells. (I) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. (J) Bar graphs showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. Data are the mean ± SD of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: ANOVA, analysis of variance; Apo, apocynin; DMSO, dimethylsulfoxide; NAC, N-acetyl-L-cysteine; PLB, plumbagin; ROS, reactive oxygen species; SD, standard deviation; 7-AAD, 7-aminoactinomycin D; PE, phycoerythrin.
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f10-dddt-9-1511: ROS plays a role in PLB-induced apoptosis and autophagy in PC-3 and DU145 cells.Notes: (A) PLB stimulating the generation of ROS in PC-3 and DU145 cells. Cells were treated with PLB at 0.1 μM, 1 μM, or 5 μM for 24 hours. The bar graphs showing the intracellular level of ROS in PC-3 and DU145 cells. (B) Bar graphs showing the time course of PLB-induced generation of intracellular ROS in PC-3 and DU145 cells. (C) Effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (D) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (E) Representative flow cytometric dot plots showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells. (F) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells determined by flow cytometry. (G) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (H) Bar graphs showing the effects of CDDO-Me on basal and PBL-induced apoptosis in PC-3 and DU145 cells. (I) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. (J) Bar graphs showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. Data are the mean ± SD of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: ANOVA, analysis of variance; Apo, apocynin; DMSO, dimethylsulfoxide; NAC, N-acetyl-L-cysteine; PLB, plumbagin; ROS, reactive oxygen species; SD, standard deviation; 7-AAD, 7-aminoactinomycin D; PE, phycoerythrin.

Mentions: To determine the effect of PLB on intracellular ROS production in PC-3 and DU145 cells, cells were treated with PLB at 0.1 μM, 1 μM, and 5 μM for 24 hours. In PC-3 cells, the intracellular level of ROS was significantly increased in a concentration-dependent manner (Figure 10A). There was a twofold increase in intracellular ROS level at 5 μM of PLB compared to the control PC-3 cells (P<0.01). In DU145 cells, there was a significant increase in intracellular ROS production after PLB treatment at 0.1 μM, 1 μM, and 5 μM for 24 hours (P<0.01; Figure 10A). In addition, co-incubation with 0.1 μM Apo, an NADPH oxidase inhibitor, significantly suppressed intracellular ROS production induced by PLB treatment in both PC-3 and DU145 cells (P<0.05; Figure 10B). In separate experiments, the ROS-inducing effect of PLB on PC-3 and DU145 cells was examined from 0 to 72 hours. There was a time-dependent increase in intracellular ROS production with 5 μM PLB treatment in both cell lines (Figure 10B). Incubation of PC-3 cells with PLB for 12 hours, 24 hours, and 72 hours increased intracellular ROS level by 20.9%, 32.5% and 41.2%, respectively. In DU145 cells, 5 μM PLB resulted in 62.4%, 42.1%, 75.4%, 54.6%, 96.0%, 83.7%, and 95.0% increase for the treatment of 2 hours, 4 hours, 8 hours, 12 hours, 24 hours, 48 hours, and 72 hours (Figure 10B).


Induction of apoptosis and autophagy via sirtuin1- and PI3K/Akt/mTOR-mediated pathways by plumbagin in human prostate cancer cells.

Zhou ZW, Li XX, He ZX, Pan ST, Yang Y, Zhang X, Chow K, Yang T, Qiu JX, Zhou Q, Tan J, Wang D, Zhou SF - Drug Des Devel Ther (2015)

ROS plays a role in PLB-induced apoptosis and autophagy in PC-3 and DU145 cells.Notes: (A) PLB stimulating the generation of ROS in PC-3 and DU145 cells. Cells were treated with PLB at 0.1 μM, 1 μM, or 5 μM for 24 hours. The bar graphs showing the intracellular level of ROS in PC-3 and DU145 cells. (B) Bar graphs showing the time course of PLB-induced generation of intracellular ROS in PC-3 and DU145 cells. (C) Effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (D) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (E) Representative flow cytometric dot plots showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells. (F) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells determined by flow cytometry. (G) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (H) Bar graphs showing the effects of CDDO-Me on basal and PBL-induced apoptosis in PC-3 and DU145 cells. (I) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. (J) Bar graphs showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. Data are the mean ± SD of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: ANOVA, analysis of variance; Apo, apocynin; DMSO, dimethylsulfoxide; NAC, N-acetyl-L-cysteine; PLB, plumbagin; ROS, reactive oxygen species; SD, standard deviation; 7-AAD, 7-aminoactinomycin D; PE, phycoerythrin.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4366042&req=5

f10-dddt-9-1511: ROS plays a role in PLB-induced apoptosis and autophagy in PC-3 and DU145 cells.Notes: (A) PLB stimulating the generation of ROS in PC-3 and DU145 cells. Cells were treated with PLB at 0.1 μM, 1 μM, or 5 μM for 24 hours. The bar graphs showing the intracellular level of ROS in PC-3 and DU145 cells. (B) Bar graphs showing the time course of PLB-induced generation of intracellular ROS in PC-3 and DU145 cells. (C) Effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (D) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (E) Representative flow cytometric dot plots showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells. (F) Bar graphs showing the effects of Apo and NAC on basal and PLB-induced autophagy in PC-3 and DU145 cells determined by flow cytometry. (G) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced apoptosis in PC-3 and DU145 cells. (H) Bar graphs showing the effects of CDDO-Me on basal and PBL-induced apoptosis in PC-3 and DU145 cells. (I) Representative flow cytometric dot plots showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. (J) Bar graphs showing the effects of CDDO-Me on basal and PLB-induced autophagy in PC-3 and DU145 cells. Data are the mean ± SD of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: ANOVA, analysis of variance; Apo, apocynin; DMSO, dimethylsulfoxide; NAC, N-acetyl-L-cysteine; PLB, plumbagin; ROS, reactive oxygen species; SD, standard deviation; 7-AAD, 7-aminoactinomycin D; PE, phycoerythrin.
Mentions: To determine the effect of PLB on intracellular ROS production in PC-3 and DU145 cells, cells were treated with PLB at 0.1 μM, 1 μM, and 5 μM for 24 hours. In PC-3 cells, the intracellular level of ROS was significantly increased in a concentration-dependent manner (Figure 10A). There was a twofold increase in intracellular ROS level at 5 μM of PLB compared to the control PC-3 cells (P<0.01). In DU145 cells, there was a significant increase in intracellular ROS production after PLB treatment at 0.1 μM, 1 μM, and 5 μM for 24 hours (P<0.01; Figure 10A). In addition, co-incubation with 0.1 μM Apo, an NADPH oxidase inhibitor, significantly suppressed intracellular ROS production induced by PLB treatment in both PC-3 and DU145 cells (P<0.05; Figure 10B). In separate experiments, the ROS-inducing effect of PLB on PC-3 and DU145 cells was examined from 0 to 72 hours. There was a time-dependent increase in intracellular ROS production with 5 μM PLB treatment in both cell lines (Figure 10B). Incubation of PC-3 cells with PLB for 12 hours, 24 hours, and 72 hours increased intracellular ROS level by 20.9%, 32.5% and 41.2%, respectively. In DU145 cells, 5 μM PLB resulted in 62.4%, 42.1%, 75.4%, 54.6%, 96.0%, 83.7%, and 95.0% increase for the treatment of 2 hours, 4 hours, 8 hours, 12 hours, 24 hours, 48 hours, and 72 hours (Figure 10B).

Bottom Line: Our study has shown that PLB had potent pro-apoptotic and pro-autophagic effects on PC-3 and DU145 cells.In addition, PLB downregulated pre-B cell colony-enhancing factor/visfatin, and the inhibition of pre-B cell colony-enhancing factor/visfatin significantly enhanced basal and PLB-induced apoptosis and autophagy in both cell lines.Moreover, reduction of intracellular reactive oxygen species (ROS) level attenuated the apoptosis- and autophagy-inducing effects of PLB on both PC-3 and DU145 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA ; Guizhou Provincial Key Laboratory for Regenerative Medicine, Stem Cell and Tissue Engineering Research Center and Sino-US Joint Laboratory for Medical Sciences, Guizhou Medical University, Guiyang, Guizhou, People's Republic of China.

ABSTRACT
Plumbagin (PLB) has been shown to have anticancer activities in animal models, but the role of PLB in prostate cancer treatment is unclear. This study aimed to investigate the effects of PLB on apoptosis and autophagy and the underlying mechanisms in human prostate cancer cell lines PC-3 and DU145. Our study has shown that PLB had potent pro-apoptotic and pro-autophagic effects on PC-3 and DU145 cells. PLB induced mitochondria-mediated apoptosis and autophagy in concentration- and time-dependent manners in both PC-3 and DU145 cells. PLB induced inhibition of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) and p38 mitogen-activated protein kinase (MAPK) pathways and activation of 5'-AMP-dependent kinase (AMPK) as indicated by their altered phosphorylation, contributing to the pro-autophagic activity of PLB. Modulation of autophagy altered basal and PLB-induced apoptosis in both cell lines. Furthermore, PLB downregulated sirtuin 1 (Sirt1), and inhibition of Sirt1 enhanced autophagy, whereas the induction of Sirt1 abolished PLB-induced autophagy in PC-3 and DU145 cells. In addition, PLB downregulated pre-B cell colony-enhancing factor/visfatin, and the inhibition of pre-B cell colony-enhancing factor/visfatin significantly enhanced basal and PLB-induced apoptosis and autophagy in both cell lines. Moreover, reduction of intracellular reactive oxygen species (ROS) level attenuated the apoptosis- and autophagy-inducing effects of PLB on both PC-3 and DU145 cells. These findings indicate that PLB promotes apoptosis and autophagy in prostate cancer cells via Sirt1- and PI3K/Akt/mTOR-mediated pathways with contribution from AMPK-, p38 MAPK-, visfatin-, and ROS-associated pathways.

No MeSH data available.


Related in: MedlinePlus