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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.

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BrMC-induced ovarian cancer cell apoptosis involved in Bim expression. (A and B) The expression levels of Bcl-2 family proteins in chemosensitive and -resistant ovarian cancer cells were examined by the time-dependent analysis and western blot analysis. A2780 and A2780/DDP cells were treated at the indicated concentrations with BrMC (5 and 10 µM) for 24 h. β-Actin was used as a control. (C) Under the same conditions as A, cells were transiently transfected with a control non-specific siRNA or Bim-specific siRNA for 48 h, and treated with or without BrMC for 24 h. Cell lysates were obtained and assayed for Bim, caspase-9 and cleaved caspase-3 by western blot. (D) Effects of Bim on the apoptotic rate in A2780 and A2780/DDP cells. All data were depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01, vs. treatment with siRNA alone; #P<0.05, vs. treatment with BrMC (5 µmol/l) and Bim siRNA. BrMC, 8-bromo-7-methoxychrysin; Bcl-2, B-cell lymphoma 2; DDP, cisplatin; siRNA, small interfering RNA; PUMA, p53-upregulated modulator of apoptosis; Bax, Bcl-2 associated X protein; Bim, Bcl2-interacting mediator of cell death; Bcl-XL, B cell lymphoma extra large; Casp, caspase; Con, control non-specific siRNA; BIM, bim-specific siRNA.
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f2-mmr-12-04-5100: BrMC-induced ovarian cancer cell apoptosis involved in Bim expression. (A and B) The expression levels of Bcl-2 family proteins in chemosensitive and -resistant ovarian cancer cells were examined by the time-dependent analysis and western blot analysis. A2780 and A2780/DDP cells were treated at the indicated concentrations with BrMC (5 and 10 µM) for 24 h. β-Actin was used as a control. (C) Under the same conditions as A, cells were transiently transfected with a control non-specific siRNA or Bim-specific siRNA for 48 h, and treated with or without BrMC for 24 h. Cell lysates were obtained and assayed for Bim, caspase-9 and cleaved caspase-3 by western blot. (D) Effects of Bim on the apoptotic rate in A2780 and A2780/DDP cells. All data were depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01, vs. treatment with siRNA alone; #P<0.05, vs. treatment with BrMC (5 µmol/l) and Bim siRNA. BrMC, 8-bromo-7-methoxychrysin; Bcl-2, B-cell lymphoma 2; DDP, cisplatin; siRNA, small interfering RNA; PUMA, p53-upregulated modulator of apoptosis; Bax, Bcl-2 associated X protein; Bim, Bcl2-interacting mediator of cell death; Bcl-XL, B cell lymphoma extra large; Casp, caspase; Con, control non-specific siRNA; BIM, bim-specific siRNA.

Mentions: Mitochondrial dysfunction has an important role in apoptosis in ovarian cells (32). Numerous studies have reported that it regulated gene expression of Bcl-2-family proteins involved in chrysin-induced apoptosis (32) and that the BH3-only proteins were necessary for chrysin-induced apoptosis in cancer cells (33). However, it remains elusive whether BH3 proteins have the same function in ovarian cancer cells following BrMC treatment. Therefore, the present study investigated the expression of Bcl-2-family proteins in cisplatin-sensitive and -resistant cells following BrMC treatment. The levels of pro-apoptotic proteins, including Bax, PUMA and NOXA, showed a slight change in A2780 cells at various time-points following BrMC treatment (Fig. 2A). No major changes were recognized in anti-apoptotic Bcl-2 and Bcl-XL proteins. However, the expression levels of Bim showed a marked increase following BrMC treatment, providing evidence that Bim was involved in apoptotic cell death in ovarian cancer cells treated with BrMC. BrMC induced Bim was expressed in the same manner in A2780/DDP cells (Fig. 2B). These results implied that Bim had an important role in BrMC-induced apoptosis in ovarian cancer cells.


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-induced ovarian cancer cell apoptosis involved in Bim expression. (A and B) The expression levels of Bcl-2 family proteins in chemosensitive and -resistant ovarian cancer cells were examined by the time-dependent analysis and western blot analysis. A2780 and A2780/DDP cells were treated at the indicated concentrations with BrMC (5 and 10 µM) for 24 h. β-Actin was used as a control. (C) Under the same conditions as A, cells were transiently transfected with a control non-specific siRNA or Bim-specific siRNA for 48 h, and treated with or without BrMC for 24 h. Cell lysates were obtained and assayed for Bim, caspase-9 and cleaved caspase-3 by western blot. (D) Effects of Bim on the apoptotic rate in A2780 and A2780/DDP cells. All data were depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01, vs. treatment with siRNA alone; #P<0.05, vs. treatment with BrMC (5 µmol/l) and Bim siRNA. BrMC, 8-bromo-7-methoxychrysin; Bcl-2, B-cell lymphoma 2; DDP, cisplatin; siRNA, small interfering RNA; PUMA, p53-upregulated modulator of apoptosis; Bax, Bcl-2 associated X protein; Bim, Bcl2-interacting mediator of cell death; Bcl-XL, B cell lymphoma extra large; Casp, caspase; Con, control non-specific siRNA; BIM, bim-specific siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2-mmr-12-04-5100: BrMC-induced ovarian cancer cell apoptosis involved in Bim expression. (A and B) The expression levels of Bcl-2 family proteins in chemosensitive and -resistant ovarian cancer cells were examined by the time-dependent analysis and western blot analysis. A2780 and A2780/DDP cells were treated at the indicated concentrations with BrMC (5 and 10 µM) for 24 h. β-Actin was used as a control. (C) Under the same conditions as A, cells were transiently transfected with a control non-specific siRNA or Bim-specific siRNA for 48 h, and treated with or without BrMC for 24 h. Cell lysates were obtained and assayed for Bim, caspase-9 and cleaved caspase-3 by western blot. (D) Effects of Bim on the apoptotic rate in A2780 and A2780/DDP cells. All data were depicted graphically as the mean ± standard error of the mean for at least three independent experiments. *P<0.05, **P<0.01, vs. treatment with siRNA alone; #P<0.05, vs. treatment with BrMC (5 µmol/l) and Bim siRNA. BrMC, 8-bromo-7-methoxychrysin; Bcl-2, B-cell lymphoma 2; DDP, cisplatin; siRNA, small interfering RNA; PUMA, p53-upregulated modulator of apoptosis; Bax, Bcl-2 associated X protein; Bim, Bcl2-interacting mediator of cell death; Bcl-XL, B cell lymphoma extra large; Casp, caspase; Con, control non-specific siRNA; BIM, bim-specific siRNA.
Mentions: Mitochondrial dysfunction has an important role in apoptosis in ovarian cells (32). Numerous studies have reported that it regulated gene expression of Bcl-2-family proteins involved in chrysin-induced apoptosis (32) and that the BH3-only proteins were necessary for chrysin-induced apoptosis in cancer cells (33). However, it remains elusive whether BH3 proteins have the same function in ovarian cancer cells following BrMC treatment. Therefore, the present study investigated the expression of Bcl-2-family proteins in cisplatin-sensitive and -resistant cells following BrMC treatment. The levels of pro-apoptotic proteins, including Bax, PUMA and NOXA, showed a slight change in A2780 cells at various time-points following BrMC treatment (Fig. 2A). No major changes were recognized in anti-apoptotic Bcl-2 and Bcl-XL proteins. However, the expression levels of Bim showed a marked increase following BrMC treatment, providing evidence that Bim was involved in apoptotic cell death in ovarian cancer cells treated with BrMC. BrMC induced Bim was expressed in the same manner in A2780/DDP cells (Fig. 2B). These results implied that Bim had an important role in BrMC-induced apoptosis in ovarian cancer cells.

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