Limits...
Chemopreventive Activity of Ferulago angulate against Breast Tumor in Rats and the Apoptotic Effect of Polycerasoidin in MCF7 Cells: A Bioassay-Guided Approach.

Karimian H, Fadaeinasab M, Zorofchian Moghadamtousi S, Hajrezaei M, Razavi M, Safi SZ, Ameen Abdulla M, Mohd Ali H, Ibrahim Noordin M - PLoS ONE (2015)

Bottom Line: Ferulago angulata leaf hexane extract (FALHE) was found to be a potent inducer of MCF7 cell apoptosis.In addition, flow cytometric analysis demonstrated that the treated MCF7 cells were arrested at the G1 phase, and this was associated with the up-regulation of p21 and p27 at both the mRNA and protein levels.The results of the present study reinforce further investigations scrutinizing the promising potential of the F. angulata chemical constituents as breast cancer chemopreventive agents.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

ABSTRACT
Ferulago angulata leaf hexane extract (FALHE) was found to be a potent inducer of MCF7 cell apoptosis. The aims of the present study were to investigate the in vivo chemopreventive effect of FALHE in rats, to identify the contributing anticancer compound in FALHE and to determine its potential mechanism of action against MCF7 cells. Thirty rats harboring LA7-induced breast tumors were divided into five groups: tumor control, low-dose FALHE, high-dose FALHE, treatment control (tamoxifen) and normal control. Breast tissues were then subjected to histopathological and immunohistochemical analyses. A bioassay-guided investigation on FALHE was performed to identify the cytotoxic compound and its mechanism of action through flow cytometry, real-time qPCR and western blotting analyses. An in vivo study showed that FALHE suppressed the expression of the tumor markers PCNA and Ki67. The tumor size was reduced from 2031 ± 281 mm3 to 432 ± 201 mm3 after FALHE treatment. FALHE administration induced apoptosis in breast tumor cells, and this was confirmed by high expression levels of Bax, p53 and caspase 3. Cell cycle arrest was suggested by the expression of p21 and p27. The in vitro experimental results resulted in the isolation of polycerasoidin as a bioactive ingredient of FALHE with an IC50 value of 3.16 ± 0.31 μg/ml against MCF7 cells. Polycerasoidin induced mitochondrial-dependent apoptosis in breast cancer cells via caspase activation and changes in the mRNA and protein expression of Bax and Bcl-2. In addition, flow cytometric analysis demonstrated that the treated MCF7 cells were arrested at the G1 phase, and this was associated with the up-regulation of p21 and p27 at both the mRNA and protein levels. The results of the present study reinforce further investigations scrutinizing the promising potential of the F. angulata chemical constituents as breast cancer chemopreventive agents.

No MeSH data available.


Related in: MedlinePlus

Western blotting analysis of apoptosis and cell cycle-related proteins normalized against the positive control β-actin.Polycerasoidin effects on protein expression of MCF7 treated cells. MCF7 cells were treated with IC50 concentration of polycerasoidin for 12, 24 and 48 h. Total protein were extracted from the cells and western blot was performed. The results showed a downregulation of Bcl-2 and upregulation of Bax, caspase 9, caspase 7, p21 and p27 in polycerasoidin-treated MCF7 cells after 12, 24 and 48 h.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4440818&req=5

pone.0127434.g012: Western blotting analysis of apoptosis and cell cycle-related proteins normalized against the positive control β-actin.Polycerasoidin effects on protein expression of MCF7 treated cells. MCF7 cells were treated with IC50 concentration of polycerasoidin for 12, 24 and 48 h. Total protein were extracted from the cells and western blot was performed. The results showed a downregulation of Bcl-2 and upregulation of Bax, caspase 9, caspase 7, p21 and p27 in polycerasoidin-treated MCF7 cells after 12, 24 and 48 h.

Mentions: To confirm the induction of apoptosis by polycerasoidin, the activities of caspase 9 and 7 in the treated MCF7 cells were examined through bioluminescent analysis (Fig 10). In addition, the expression of these two cysteinyl aspartate proteinases was investigated at the mRNA and protein levels. As illustrated in Fig 11, both caspase 9 and caspase 7 were activated significantly after 12 h of treatment with polycerasoidin. Thus, the mRNA and protein expression of these caspases was examined after 12, 24 and 48 h of treatment. The mRNA expression of caspase 9 in MCF7 cells treated with an IC50 dose of polycerasoidin was elevated by 2.06 ± 0.39-fold at 12 h and reached 4.6 ± 0.64-fold and 6.5 ± 0.94-fold higher levels at 24 and 48 h, respectively. Similarly, the caspase 7 mRNA expression also increased time-dependently from 0.41 ± 0.31-fold at 12 h to 2.85 ± 0.88-fold at 48 h (Fig 11). Western blotting analysis also confirmed this upregulation (Fig 12), which strongly demonstrated the induction of apoptosis by polycerasoidin and shed new light on the involvement of the apoptosis pathway.


Chemopreventive Activity of Ferulago angulate against Breast Tumor in Rats and the Apoptotic Effect of Polycerasoidin in MCF7 Cells: A Bioassay-Guided Approach.

Karimian H, Fadaeinasab M, Zorofchian Moghadamtousi S, Hajrezaei M, Razavi M, Safi SZ, Ameen Abdulla M, Mohd Ali H, Ibrahim Noordin M - PLoS ONE (2015)

Western blotting analysis of apoptosis and cell cycle-related proteins normalized against the positive control β-actin.Polycerasoidin effects on protein expression of MCF7 treated cells. MCF7 cells were treated with IC50 concentration of polycerasoidin for 12, 24 and 48 h. Total protein were extracted from the cells and western blot was performed. The results showed a downregulation of Bcl-2 and upregulation of Bax, caspase 9, caspase 7, p21 and p27 in polycerasoidin-treated MCF7 cells after 12, 24 and 48 h.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440818&req=5

pone.0127434.g012: Western blotting analysis of apoptosis and cell cycle-related proteins normalized against the positive control β-actin.Polycerasoidin effects on protein expression of MCF7 treated cells. MCF7 cells were treated with IC50 concentration of polycerasoidin for 12, 24 and 48 h. Total protein were extracted from the cells and western blot was performed. The results showed a downregulation of Bcl-2 and upregulation of Bax, caspase 9, caspase 7, p21 and p27 in polycerasoidin-treated MCF7 cells after 12, 24 and 48 h.
Mentions: To confirm the induction of apoptosis by polycerasoidin, the activities of caspase 9 and 7 in the treated MCF7 cells were examined through bioluminescent analysis (Fig 10). In addition, the expression of these two cysteinyl aspartate proteinases was investigated at the mRNA and protein levels. As illustrated in Fig 11, both caspase 9 and caspase 7 were activated significantly after 12 h of treatment with polycerasoidin. Thus, the mRNA and protein expression of these caspases was examined after 12, 24 and 48 h of treatment. The mRNA expression of caspase 9 in MCF7 cells treated with an IC50 dose of polycerasoidin was elevated by 2.06 ± 0.39-fold at 12 h and reached 4.6 ± 0.64-fold and 6.5 ± 0.94-fold higher levels at 24 and 48 h, respectively. Similarly, the caspase 7 mRNA expression also increased time-dependently from 0.41 ± 0.31-fold at 12 h to 2.85 ± 0.88-fold at 48 h (Fig 11). Western blotting analysis also confirmed this upregulation (Fig 12), which strongly demonstrated the induction of apoptosis by polycerasoidin and shed new light on the involvement of the apoptosis pathway.

Bottom Line: Ferulago angulata leaf hexane extract (FALHE) was found to be a potent inducer of MCF7 cell apoptosis.In addition, flow cytometric analysis demonstrated that the treated MCF7 cells were arrested at the G1 phase, and this was associated with the up-regulation of p21 and p27 at both the mRNA and protein levels.The results of the present study reinforce further investigations scrutinizing the promising potential of the F. angulata chemical constituents as breast cancer chemopreventive agents.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

ABSTRACT
Ferulago angulata leaf hexane extract (FALHE) was found to be a potent inducer of MCF7 cell apoptosis. The aims of the present study were to investigate the in vivo chemopreventive effect of FALHE in rats, to identify the contributing anticancer compound in FALHE and to determine its potential mechanism of action against MCF7 cells. Thirty rats harboring LA7-induced breast tumors were divided into five groups: tumor control, low-dose FALHE, high-dose FALHE, treatment control (tamoxifen) and normal control. Breast tissues were then subjected to histopathological and immunohistochemical analyses. A bioassay-guided investigation on FALHE was performed to identify the cytotoxic compound and its mechanism of action through flow cytometry, real-time qPCR and western blotting analyses. An in vivo study showed that FALHE suppressed the expression of the tumor markers PCNA and Ki67. The tumor size was reduced from 2031 ± 281 mm3 to 432 ± 201 mm3 after FALHE treatment. FALHE administration induced apoptosis in breast tumor cells, and this was confirmed by high expression levels of Bax, p53 and caspase 3. Cell cycle arrest was suggested by the expression of p21 and p27. The in vitro experimental results resulted in the isolation of polycerasoidin as a bioactive ingredient of FALHE with an IC50 value of 3.16 ± 0.31 μg/ml against MCF7 cells. Polycerasoidin induced mitochondrial-dependent apoptosis in breast cancer cells via caspase activation and changes in the mRNA and protein expression of Bax and Bcl-2. In addition, flow cytometric analysis demonstrated that the treated MCF7 cells were arrested at the G1 phase, and this was associated with the up-regulation of p21 and p27 at both the mRNA and protein levels. The results of the present study reinforce further investigations scrutinizing the promising potential of the F. angulata chemical constituents as breast cancer chemopreventive agents.

No MeSH data available.


Related in: MedlinePlus