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Antiproliferative activity and induction of apoptosis in estrogen receptor-positive and negative human breast carcinoma cell lines by Gmelina asiatica roots.

Balijepalli MK, Tandra S, Pichika MR - Pharmacognosy Res (2010)

Bottom Line: The EGAR contain lignans and flavonoids.The antiproliferative activity of the extract is attributed to the presence of these secondary metabolites.The results suggest the efficacy of G. asiatica roots as antiproliferative agents on human breast cancer cells, supporting the hypothesis that plants containing lignans have beneficial effects on human breast cancer.

View Article: PubMed Central - PubMed

Affiliation: International Medical University, Kuala Lumpur, Malaysia, India.

ABSTRACT
Low risk of breast cancer has been proposed to be associated with high intake of lignans. We have reported the presence of lignans in Gmelina asiatica roots. There are no scientific reports on the antiproliferative activity of G. asiatica roots. The objective of the present study was to evaluate the effect of ethyl acetate extract from G. asiatica roots (EGAR) on estrogen receptor-positive (MCF-7) and negative (MDA-MB-231) human breast cancer cell lines. The effects of 50% inhibitory concentrations (IC(50)) of EGAR on MCF-7 and MDA-MB-231 cells were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay kit. The mode of cell death caused by EGAR was determined using dual apoptosis assay kit by observing the cells under fluorescent microscope. The quantification of apoptosis and necrosis in cells caused by EGAR was determined using cell death detection kit through ELISA. Down-regulation of the proliferative activity occurred in a clear dose-dependent response with IC(50) values of 32.9 ± 3.8 μg/mL in MCF-7 and 19.9 ± 2.3 μg/mL in MDA-MB-231 cell lines. Treatment of breast cancer cells with EGAR resulted in significant apoptosis. The EGAR contain lignans and flavonoids. The antiproliferative activity of the extract is attributed to the presence of these secondary metabolites. The results suggest the efficacy of G. asiatica roots as antiproliferative agents on human breast cancer cells, supporting the hypothesis that plants containing lignans have beneficial effects on human breast cancer.

No MeSH data available.


Related in: MedlinePlus

Merged photomicrograph of stained MDA-MB-231 cell lines after 6 h treatment with 20 μg/mL of ethylacetate extract from G. asiatica roots. Green nucleus surrounded by red border indicating the apoptoic MDA-MB-231 cells
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Figure 5: Merged photomicrograph of stained MDA-MB-231 cell lines after 6 h treatment with 20 μg/mL of ethylacetate extract from G. asiatica roots. Green nucleus surrounded by red border indicating the apoptoic MDA-MB-231 cells

Mentions: In principle, a reduction of cell growth can reflect either a decreased proliferation rate or an enhanced cell death by either necrosis or apoptosis or a combination of these two mechanisms. We also investigated whether the extracts can induce apoptosis by monitoring the two important apoptosis markers: phosphatidylserine (PS) translocation on cell membrane (identified by sulforhodamine 101-annexin V)[13] and caspase-3 activation (identified by NucView™ 488 caspase-3 substrate).[14] The morphological changes were inspected by microscopy. Some cells were beginning to detach from the plate and become rounded after 3 hours of treatment by EGAR (30 μg/mL for MCF-7 and 20 μg/mL for MDA-MB-231). Because loss of adhesion to the culture dishes of tumoral epithelial cells has been described as an apoptosis-related event,[15] we examined the morphological apoptotic changes on slides under bright field after 3 hours and 6 hours. In contrast to good spread cells in the negative control, a morphological change with cell shrinkage was detected in cell lines treated with EGAR extract. Necrosis was less prominent than apoptosis. In Figures 1a–e, the representative photomicrographs of MDA-MB-231 cells after treatment with 20 μg/mL EGAR extract in comparison to solvent-treated control are shown.


Antiproliferative activity and induction of apoptosis in estrogen receptor-positive and negative human breast carcinoma cell lines by Gmelina asiatica roots.

Balijepalli MK, Tandra S, Pichika MR - Pharmacognosy Res (2010)

Merged photomicrograph of stained MDA-MB-231 cell lines after 6 h treatment with 20 μg/mL of ethylacetate extract from G. asiatica roots. Green nucleus surrounded by red border indicating the apoptoic MDA-MB-231 cells
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3140107&req=5

Figure 5: Merged photomicrograph of stained MDA-MB-231 cell lines after 6 h treatment with 20 μg/mL of ethylacetate extract from G. asiatica roots. Green nucleus surrounded by red border indicating the apoptoic MDA-MB-231 cells
Mentions: In principle, a reduction of cell growth can reflect either a decreased proliferation rate or an enhanced cell death by either necrosis or apoptosis or a combination of these two mechanisms. We also investigated whether the extracts can induce apoptosis by monitoring the two important apoptosis markers: phosphatidylserine (PS) translocation on cell membrane (identified by sulforhodamine 101-annexin V)[13] and caspase-3 activation (identified by NucView™ 488 caspase-3 substrate).[14] The morphological changes were inspected by microscopy. Some cells were beginning to detach from the plate and become rounded after 3 hours of treatment by EGAR (30 μg/mL for MCF-7 and 20 μg/mL for MDA-MB-231). Because loss of adhesion to the culture dishes of tumoral epithelial cells has been described as an apoptosis-related event,[15] we examined the morphological apoptotic changes on slides under bright field after 3 hours and 6 hours. In contrast to good spread cells in the negative control, a morphological change with cell shrinkage was detected in cell lines treated with EGAR extract. Necrosis was less prominent than apoptosis. In Figures 1a–e, the representative photomicrographs of MDA-MB-231 cells after treatment with 20 μg/mL EGAR extract in comparison to solvent-treated control are shown.

Bottom Line: The EGAR contain lignans and flavonoids.The antiproliferative activity of the extract is attributed to the presence of these secondary metabolites.The results suggest the efficacy of G. asiatica roots as antiproliferative agents on human breast cancer cells, supporting the hypothesis that plants containing lignans have beneficial effects on human breast cancer.

View Article: PubMed Central - PubMed

Affiliation: International Medical University, Kuala Lumpur, Malaysia, India.

ABSTRACT
Low risk of breast cancer has been proposed to be associated with high intake of lignans. We have reported the presence of lignans in Gmelina asiatica roots. There are no scientific reports on the antiproliferative activity of G. asiatica roots. The objective of the present study was to evaluate the effect of ethyl acetate extract from G. asiatica roots (EGAR) on estrogen receptor-positive (MCF-7) and negative (MDA-MB-231) human breast cancer cell lines. The effects of 50% inhibitory concentrations (IC(50)) of EGAR on MCF-7 and MDA-MB-231 cells were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay kit. The mode of cell death caused by EGAR was determined using dual apoptosis assay kit by observing the cells under fluorescent microscope. The quantification of apoptosis and necrosis in cells caused by EGAR was determined using cell death detection kit through ELISA. Down-regulation of the proliferative activity occurred in a clear dose-dependent response with IC(50) values of 32.9 ± 3.8 μg/mL in MCF-7 and 19.9 ± 2.3 μg/mL in MDA-MB-231 cell lines. Treatment of breast cancer cells with EGAR resulted in significant apoptosis. The EGAR contain lignans and flavonoids. The antiproliferative activity of the extract is attributed to the presence of these secondary metabolites. The results suggest the efficacy of G. asiatica roots as antiproliferative agents on human breast cancer cells, supporting the hypothesis that plants containing lignans have beneficial effects on human breast cancer.

No MeSH data available.


Related in: MedlinePlus