Limits...
8‑bromo‑7‑methoxychrysin induces apoptosis by regulating Akt/FOXO3a pathway in cisplatin‑sensitive and resistant ovarian cancer cells.

Ding Q, Chen Y, Zhang Q, Guo Y, Huang Z, Dai L, Cao S - Mol Med Rep (2015)

Bottom Line: 8‑bromo‑7‑methoxychrysin (BrMC), a novel chrysin analog, was reported to have anti‑cancer activities.BrMC‑induced apoptotic cell death occurred mainly by the activation of Akt, which was accompanied by the overexpression of transcription factor FOXO3a, with a concomitant increase in the expression levels of Bim.Silencing Bim expression by using small interfering RNA, attenuated the induction of apoptosis by BrMC treatment.

View Article: PubMed Central - PubMed

Affiliation: Hunan University of Traditional Chinese Medicine, Changsha, Hunan 410208, P.R.China.

ABSTRACT
8‑bromo‑7‑methoxychrysin (BrMC), a novel chrysin analog, was reported to have anti‑cancer activities. The aim of the present study was to investigate the molecular mechanism of 8‑bromo‑7‑methoxychrysin (BrMC)‑induced apoptosis via the Akt/forkhead box O3a (FOXO3a) pathway in cisplatin (DDP)‑sensitive and ‑resistant ovarian cancer cells. The human ovarian cancer cell lines A2780 and A2780/DDP were cultured in vitro. Various molecular techniques were used to assess the expression of FOXO3a and B cell lymphoma 2 (Bcl‑2)‑interacting mediator of cell death (Bim) in cisplatin‑sensitive and ‑resistant ovarian cancer cells. Different concentrations of BrMC induced apoptosis in cisplatin‑sensitive and ‑resistant ovarian cancer cells. BrMC‑induced apoptotic cell death occurred mainly by the activation of Akt, which was accompanied by the overexpression of transcription factor FOXO3a, with a concomitant increase in the expression levels of Bim. Silencing Bim expression by using small interfering RNA, attenuated the induction of apoptosis by BrMC treatment. The results indicated that BrMC‑induced apoptosis in cisplatin‑sensitive and ‑resistant ovarian cancer cells may occur via the regulation of Akt/FOXO3a, leading to Bim transcription.

Show MeSH

Related in: MedlinePlus

BrMC induces apoptosis in cisplatin-sensitive and -resistant human ovarian cells. (A and B) Effect of cisplatin on apoptosis in ovarian cancer cell lines in a dose-dependent manner. Cells were treated with the indicated concentrations of cisplatin for 48 h. ELISA was used to determine histone/DNA fragmentation. Data are presented as the mean ± SD (n=4). *P<0.05, **P<0.01 vs. 0.1% DMSO; #P<0.05, ##P<0.01 vs. treatment with 2.5 µmol/l BrMC. (C and D) Effect of BrMC on apoptotic death in cisplatin-sensitive and -resistant cells. Apoptosis was detected using propidium iodide and flow cytometric analysis. Cells were cultured in the presence or absence of BrMC (2.5 µM) for different time periods, and apoptosis was measured. All data are depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01. (E and F) Analyses of cyt c release. Following BrMC treatment for different periods of time, cells were subjected to sub-cellular fractionation. The specific antibodies to cyt c, β-actin and Cox IV were used. Densitometric analysis of the western blots was performed and the amount of cyt c was compared with that of the loading control. Data are representative of at least three independent experiments. BrMC, 8-bromo-7-methoxychrysin; SD, standard deviation; DMSO, dimethyl sulfoxide; DDP, cisplatin; cyt c, cytochrome c; Cox IV, cytochrome c oxidase IV; Cyto, cytosol; Mito, mitochondria.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4581817&req=5

f1-mmr-12-04-5100: BrMC induces apoptosis in cisplatin-sensitive and -resistant human ovarian cells. (A and B) Effect of cisplatin on apoptosis in ovarian cancer cell lines in a dose-dependent manner. Cells were treated with the indicated concentrations of cisplatin for 48 h. ELISA was used to determine histone/DNA fragmentation. Data are presented as the mean ± SD (n=4). *P<0.05, **P<0.01 vs. 0.1% DMSO; #P<0.05, ##P<0.01 vs. treatment with 2.5 µmol/l BrMC. (C and D) Effect of BrMC on apoptotic death in cisplatin-sensitive and -resistant cells. Apoptosis was detected using propidium iodide and flow cytometric analysis. Cells were cultured in the presence or absence of BrMC (2.5 µM) for different time periods, and apoptosis was measured. All data are depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01. (E and F) Analyses of cyt c release. Following BrMC treatment for different periods of time, cells were subjected to sub-cellular fractionation. The specific antibodies to cyt c, β-actin and Cox IV were used. Densitometric analysis of the western blots was performed and the amount of cyt c was compared with that of the loading control. Data are representative of at least three independent experiments. BrMC, 8-bromo-7-methoxychrysin; SD, standard deviation; DMSO, dimethyl sulfoxide; DDP, cisplatin; cyt c, cytochrome c; Cox IV, cytochrome c oxidase IV; Cyto, cytosol; Mito, mitochondria.

Mentions: The growth inhibitory effect of cisplatin was determined in various cisplatin-sensitive (A2780) and -resistant (A2780/DDP) human ovarian cell lines. Cell viability was detected by the MTT assay after 48 h of treatment with cisplatin. As shown in Fig. 1A, cisplatin caused a dose-dependent reduction of cell viability in A2780 cells, while A2780/DDP cells exhibited resistance to cisplatin. To examine cisplatin-induced apoptosis, the cells were treated with cisplatin and apoptosis was confirmed by a histone/DNA fragmentation ELISA assay at various concentrations. Cisplatin effectively induced apoptosis in cisplatin-sensitive ovarian cancer cells, but not in cisplatin-resistant cells (Fig. 1B). To investigate the effects of BrMC on apoptosis in both ovarian cancer cell lines, the ovarian cancer cells were treated with BrMC and the cell apoptosis process was analyzed by a DNA fragmentation ELISA assay. BrMC induced cell apoptosis in both ovarian cancer cell lines, regardless of their differences in chemosensitivity (Fig. 1C). Flow cytometric (FCM) analysis with PI staining further revealed that there was no difference in BrMC-induced apoptosis between cisplatin-sensitive and -resistant cells (Fig. 1D). To determine whether mitochondria had an important role in BrMC-induced cell apoptosis, the release of cytochrome c was detected by cell fractionation analysis. The results revealed that BrMC induced the release of cytochrome c in a time-dependent manner in both chemo-sensitive and -resistant cells (Fig. 1E and F), suggesting that BrMC initiated apoptotic cell death through mitochondrial dysfunction.


8‑bromo‑7‑methoxychrysin induces apoptosis by regulating Akt/FOXO3a pathway in cisplatin‑sensitive and resistant ovarian cancer cells.

Ding Q, Chen Y, Zhang Q, Guo Y, Huang Z, Dai L, Cao S - Mol Med Rep (2015)

BrMC induces apoptosis in cisplatin-sensitive and -resistant human ovarian cells. (A and B) Effect of cisplatin on apoptosis in ovarian cancer cell lines in a dose-dependent manner. Cells were treated with the indicated concentrations of cisplatin for 48 h. ELISA was used to determine histone/DNA fragmentation. Data are presented as the mean ± SD (n=4). *P<0.05, **P<0.01 vs. 0.1% DMSO; #P<0.05, ##P<0.01 vs. treatment with 2.5 µmol/l BrMC. (C and D) Effect of BrMC on apoptotic death in cisplatin-sensitive and -resistant cells. Apoptosis was detected using propidium iodide and flow cytometric analysis. Cells were cultured in the presence or absence of BrMC (2.5 µM) for different time periods, and apoptosis was measured. All data are depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01. (E and F) Analyses of cyt c release. Following BrMC treatment for different periods of time, cells were subjected to sub-cellular fractionation. The specific antibodies to cyt c, β-actin and Cox IV were used. Densitometric analysis of the western blots was performed and the amount of cyt c was compared with that of the loading control. Data are representative of at least three independent experiments. BrMC, 8-bromo-7-methoxychrysin; SD, standard deviation; DMSO, dimethyl sulfoxide; DDP, cisplatin; cyt c, cytochrome c; Cox IV, cytochrome c oxidase IV; Cyto, cytosol; Mito, mitochondria.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581817&req=5

f1-mmr-12-04-5100: BrMC induces apoptosis in cisplatin-sensitive and -resistant human ovarian cells. (A and B) Effect of cisplatin on apoptosis in ovarian cancer cell lines in a dose-dependent manner. Cells were treated with the indicated concentrations of cisplatin for 48 h. ELISA was used to determine histone/DNA fragmentation. Data are presented as the mean ± SD (n=4). *P<0.05, **P<0.01 vs. 0.1% DMSO; #P<0.05, ##P<0.01 vs. treatment with 2.5 µmol/l BrMC. (C and D) Effect of BrMC on apoptotic death in cisplatin-sensitive and -resistant cells. Apoptosis was detected using propidium iodide and flow cytometric analysis. Cells were cultured in the presence or absence of BrMC (2.5 µM) for different time periods, and apoptosis was measured. All data are depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01. (E and F) Analyses of cyt c release. Following BrMC treatment for different periods of time, cells were subjected to sub-cellular fractionation. The specific antibodies to cyt c, β-actin and Cox IV were used. Densitometric analysis of the western blots was performed and the amount of cyt c was compared with that of the loading control. Data are representative of at least three independent experiments. BrMC, 8-bromo-7-methoxychrysin; SD, standard deviation; DMSO, dimethyl sulfoxide; DDP, cisplatin; cyt c, cytochrome c; Cox IV, cytochrome c oxidase IV; Cyto, cytosol; Mito, mitochondria.
Mentions: The growth inhibitory effect of cisplatin was determined in various cisplatin-sensitive (A2780) and -resistant (A2780/DDP) human ovarian cell lines. Cell viability was detected by the MTT assay after 48 h of treatment with cisplatin. As shown in Fig. 1A, cisplatin caused a dose-dependent reduction of cell viability in A2780 cells, while A2780/DDP cells exhibited resistance to cisplatin. To examine cisplatin-induced apoptosis, the cells were treated with cisplatin and apoptosis was confirmed by a histone/DNA fragmentation ELISA assay at various concentrations. Cisplatin effectively induced apoptosis in cisplatin-sensitive ovarian cancer cells, but not in cisplatin-resistant cells (Fig. 1B). To investigate the effects of BrMC on apoptosis in both ovarian cancer cell lines, the ovarian cancer cells were treated with BrMC and the cell apoptosis process was analyzed by a DNA fragmentation ELISA assay. BrMC induced cell apoptosis in both ovarian cancer cell lines, regardless of their differences in chemosensitivity (Fig. 1C). Flow cytometric (FCM) analysis with PI staining further revealed that there was no difference in BrMC-induced apoptosis between cisplatin-sensitive and -resistant cells (Fig. 1D). To determine whether mitochondria had an important role in BrMC-induced cell apoptosis, the release of cytochrome c was detected by cell fractionation analysis. The results revealed that BrMC induced the release of cytochrome c in a time-dependent manner in both chemo-sensitive and -resistant cells (Fig. 1E and F), suggesting that BrMC initiated apoptotic cell death through mitochondrial dysfunction.

Bottom Line: 8‑bromo‑7‑methoxychrysin (BrMC), a novel chrysin analog, was reported to have anti‑cancer activities.BrMC‑induced apoptotic cell death occurred mainly by the activation of Akt, which was accompanied by the overexpression of transcription factor FOXO3a, with a concomitant increase in the expression levels of Bim.Silencing Bim expression by using small interfering RNA, attenuated the induction of apoptosis by BrMC treatment.

View Article: PubMed Central - PubMed

Affiliation: Hunan University of Traditional Chinese Medicine, Changsha, Hunan 410208, P.R.China.

ABSTRACT
8‑bromo‑7‑methoxychrysin (BrMC), a novel chrysin analog, was reported to have anti‑cancer activities. The aim of the present study was to investigate the molecular mechanism of 8‑bromo‑7‑methoxychrysin (BrMC)‑induced apoptosis via the Akt/forkhead box O3a (FOXO3a) pathway in cisplatin (DDP)‑sensitive and ‑resistant ovarian cancer cells. The human ovarian cancer cell lines A2780 and A2780/DDP were cultured in vitro. Various molecular techniques were used to assess the expression of FOXO3a and B cell lymphoma 2 (Bcl‑2)‑interacting mediator of cell death (Bim) in cisplatin‑sensitive and ‑resistant ovarian cancer cells. Different concentrations of BrMC induced apoptosis in cisplatin‑sensitive and ‑resistant ovarian cancer cells. BrMC‑induced apoptotic cell death occurred mainly by the activation of Akt, which was accompanied by the overexpression of transcription factor FOXO3a, with a concomitant increase in the expression levels of Bim. Silencing Bim expression by using small interfering RNA, attenuated the induction of apoptosis by BrMC treatment. The results indicated that BrMC‑induced apoptosis in cisplatin‑sensitive and ‑resistant ovarian cancer cells may occur via the regulation of Akt/FOXO3a, leading to Bim transcription.

Show MeSH
Related in: MedlinePlus