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Induction of apoptosis through oxidative stress-related pathways in MCF-7, human breast cancer cells, by ethyl acetate extract of Dillenia suffruticosa.

Tor YS, Yazan LS, Foo JB, Armania N, Cheah YK, Abdullah R, Imam MU, Ismail N, Ismail M - BMC Complement Altern Med (2014)

Bottom Line: In this study, the anticancer effect of ethyl acetate extract of D. suffruticosa (EADs) was examined on human breast adenocarcinoma cell line MCF-7 and the molecular pathway involved was elucidated.EADs induced non-phase specific cell cycle arrest at different concentration and time point.It is suggested that EADs induced apoptosis in MCF-7 cells by modulating numerous genes which are involved in oxidative stress pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia. latifahsy@upm.edu.my.

ABSTRACT

Background: Breast cancer is one of the most dreading types of cancer among women. Herbal medicine has becoming a potential source of treatment for breast cancer. Herbal plant Dillenia suffruticosa (Griff) Martelli under the family Dilleniaceae has been traditionally used to treat cancerous growth. In this study, the anticancer effect of ethyl acetate extract of D. suffruticosa (EADs) was examined on human breast adenocarcinoma cell line MCF-7 and the molecular pathway involved was elucidated.

Methods: EADs was obtained from the root of D. suffruticosa by using sequential solvent extraction. Cytotoxicity was determined by using MTT assay, mode of cell death by cell cycle analysis and apoptosis induction by Annexin-FITC/PI assay. Morphology changes in cells were observed under inverted light microscope. Involvement of selected genes in the oxidative stress-mediated signaling pathway was explored using multiplex gene expression analysis.

Results: The treatment of EADs caused cytotoxicity to MCF-7 cells in a dose- and time-dependent manner at 24, 48 and 72 hours with IC50 of 76 ± 2.3, 58 ± 0.7 and 39 ± 3.6 μg/mL, respectively. The IC50 of tamoxifen-treated MCF-7 cells was 8 ± 0.5 μg/mL. Induction of apoptosis by EADs was dose- and time- dependent. EADs induced non-phase specific cell cycle arrest at different concentration and time point. The multiplex mRNA expression study indicated that EADs-induced apoptosis was accompanied by upregulation of the expression of SOD1, SOD2, NF-κB, p53, p38 MAPK, and catalase, but downregulation of Akt1.

Conclusion: It is suggested that EADs induced apoptosis in MCF-7 cells by modulating numerous genes which are involved in oxidative stress pathway. Therefore, EADs has the potential to act as an effective intervention against breast cancer cells.

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Related in: MedlinePlus

Cell cycle analysis of MCF-7 breast cancer cells treated with EADs at 24 and 48 hours. Effects of EADs on the cell cycle distribution in MCF-7 cells were analysed using flowcytometry analysis. Bar charts representing the percentage of cell populations in MCF-7 cells treated with EADS for (a) 24 hours and (b) 48 hours. DNA histogram (Figure 3c) displayed cell cycle phase distribution of control and EADs-treated cells at 24 and 48 hours. The data are presented as mean ± standard deviation of three replicates in three independent tests. An asterisk * indicates statistically significant different from untreated control (P < 0.05).
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Figure 3: Cell cycle analysis of MCF-7 breast cancer cells treated with EADs at 24 and 48 hours. Effects of EADs on the cell cycle distribution in MCF-7 cells were analysed using flowcytometry analysis. Bar charts representing the percentage of cell populations in MCF-7 cells treated with EADS for (a) 24 hours and (b) 48 hours. DNA histogram (Figure 3c) displayed cell cycle phase distribution of control and EADs-treated cells at 24 and 48 hours. The data are presented as mean ± standard deviation of three replicates in three independent tests. An asterisk * indicates statistically significant different from untreated control (P < 0.05).

Mentions: The cell cycle phase distribution of MCF-7 cells treated with EADs at 24 and 48 hours is depicted in Figure 3. The cell cycle arrest by EADs was time- and concentration-dependent. At 24 and 48 hours, an increase in cell population in G1 at 25 μg/mL of EADs was noted (P < 0.05). On the other hand, 50 μg/mL of EADs at 24 hours elevated the number of cells in S and G2/M compared to control, accompanied by a decline in G1 phase cell population (P < 0.05). Meanwhile, at 48 hours, G2/M phase cell population was ascended compared to the control following treatment with 50 μg/mL of EADs (P < 0.05). Increase in the population of cells at sub-G1 phase was observed following treatment with EADs (P < 0.05).


Induction of apoptosis through oxidative stress-related pathways in MCF-7, human breast cancer cells, by ethyl acetate extract of Dillenia suffruticosa.

Tor YS, Yazan LS, Foo JB, Armania N, Cheah YK, Abdullah R, Imam MU, Ismail N, Ismail M - BMC Complement Altern Med (2014)

Cell cycle analysis of MCF-7 breast cancer cells treated with EADs at 24 and 48 hours. Effects of EADs on the cell cycle distribution in MCF-7 cells were analysed using flowcytometry analysis. Bar charts representing the percentage of cell populations in MCF-7 cells treated with EADS for (a) 24 hours and (b) 48 hours. DNA histogram (Figure 3c) displayed cell cycle phase distribution of control and EADs-treated cells at 24 and 48 hours. The data are presented as mean ± standard deviation of three replicates in three independent tests. An asterisk * indicates statistically significant different from untreated control (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3927215&req=5

Figure 3: Cell cycle analysis of MCF-7 breast cancer cells treated with EADs at 24 and 48 hours. Effects of EADs on the cell cycle distribution in MCF-7 cells were analysed using flowcytometry analysis. Bar charts representing the percentage of cell populations in MCF-7 cells treated with EADS for (a) 24 hours and (b) 48 hours. DNA histogram (Figure 3c) displayed cell cycle phase distribution of control and EADs-treated cells at 24 and 48 hours. The data are presented as mean ± standard deviation of three replicates in three independent tests. An asterisk * indicates statistically significant different from untreated control (P < 0.05).
Mentions: The cell cycle phase distribution of MCF-7 cells treated with EADs at 24 and 48 hours is depicted in Figure 3. The cell cycle arrest by EADs was time- and concentration-dependent. At 24 and 48 hours, an increase in cell population in G1 at 25 μg/mL of EADs was noted (P < 0.05). On the other hand, 50 μg/mL of EADs at 24 hours elevated the number of cells in S and G2/M compared to control, accompanied by a decline in G1 phase cell population (P < 0.05). Meanwhile, at 48 hours, G2/M phase cell population was ascended compared to the control following treatment with 50 μg/mL of EADs (P < 0.05). Increase in the population of cells at sub-G1 phase was observed following treatment with EADs (P < 0.05).

Bottom Line: In this study, the anticancer effect of ethyl acetate extract of D. suffruticosa (EADs) was examined on human breast adenocarcinoma cell line MCF-7 and the molecular pathway involved was elucidated.EADs induced non-phase specific cell cycle arrest at different concentration and time point.It is suggested that EADs induced apoptosis in MCF-7 cells by modulating numerous genes which are involved in oxidative stress pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia. latifahsy@upm.edu.my.

ABSTRACT

Background: Breast cancer is one of the most dreading types of cancer among women. Herbal medicine has becoming a potential source of treatment for breast cancer. Herbal plant Dillenia suffruticosa (Griff) Martelli under the family Dilleniaceae has been traditionally used to treat cancerous growth. In this study, the anticancer effect of ethyl acetate extract of D. suffruticosa (EADs) was examined on human breast adenocarcinoma cell line MCF-7 and the molecular pathway involved was elucidated.

Methods: EADs was obtained from the root of D. suffruticosa by using sequential solvent extraction. Cytotoxicity was determined by using MTT assay, mode of cell death by cell cycle analysis and apoptosis induction by Annexin-FITC/PI assay. Morphology changes in cells were observed under inverted light microscope. Involvement of selected genes in the oxidative stress-mediated signaling pathway was explored using multiplex gene expression analysis.

Results: The treatment of EADs caused cytotoxicity to MCF-7 cells in a dose- and time-dependent manner at 24, 48 and 72 hours with IC50 of 76 ± 2.3, 58 ± 0.7 and 39 ± 3.6 μg/mL, respectively. The IC50 of tamoxifen-treated MCF-7 cells was 8 ± 0.5 μg/mL. Induction of apoptosis by EADs was dose- and time- dependent. EADs induced non-phase specific cell cycle arrest at different concentration and time point. The multiplex mRNA expression study indicated that EADs-induced apoptosis was accompanied by upregulation of the expression of SOD1, SOD2, NF-κB, p53, p38 MAPK, and catalase, but downregulation of Akt1.

Conclusion: It is suggested that EADs induced apoptosis in MCF-7 cells by modulating numerous genes which are involved in oxidative stress pathway. Therefore, EADs has the potential to act as an effective intervention against breast cancer cells.

Show MeSH
Related in: MedlinePlus