Limits...
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

Time dependent apoptosis of MCF-7 and MDA-MB-231 cell lines induced by ethylacetate extract from G. asiatica roots. *Values are statistically significant at P<0.05 with respect to corresponding controls
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3140107&req=5

Figure 6: Time dependent apoptosis of MCF-7 and MDA-MB-231 cell lines induced by ethylacetate extract from G. asiatica roots. *Values are statistically significant at P<0.05 with respect to corresponding controls

Mentions: To further substantiate the antiproliferative effects of EGAR, the apoptotic cells were monitored by annexin V adherence and caspase-3 activation. In viable cells, PS is located on the cytoplasmic surface of the cell membrane; in apoptotic cells, PS is translocated from the inner to the outer leaflet of the plasma membrane, thus exposing PS. Annexin V is a Ca2+–dependent phospholipids-binding protein with high affinity for PS. The binding of sophorodamine101-annexin V probe to PS that has translocated to the outer membrane cell produces red border around the cell under fluorescent microscope using red filter [Figure 1c]. Caspase-3 (CPP32) is a cytosolic protein that normally exists as a 32-kDa inactive precursor. It is cleaved proteolytically into a heterodimer when the cell undergoes apoptosis.[16] The cleavage of NucView™488 caspase-3 substrate by activated caspase-3 stains the cell nucleus green [Figure 1d]. The induction of apoptosis by EGAR was time dependent [Figure 2]. In MDA-MB-231 cells, an elevation in apoptosis-positive cells up to 26.5% was found after 3 hours and reached 48.3% after 6 hours of exposure to 20 μg/mL EGAR. In MCF-7, there were fewer apoptotic cells, EGAR at concentration of 30 μg/mL inducing apoptosis in 28.1% and 42.9% after 3 and 6 hours of treatment, respectively. The solvent controls did not increase the spontaneous apoptotic rate in the two malignant cell lines tested.


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)

Time dependent apoptosis of MCF-7 and MDA-MB-231 cell lines induced by ethylacetate extract from G. asiatica roots. *Values are statistically significant at P<0.05 with respect to corresponding controls
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 6: Time dependent apoptosis of MCF-7 and MDA-MB-231 cell lines induced by ethylacetate extract from G. asiatica roots. *Values are statistically significant at P<0.05 with respect to corresponding controls
Mentions: To further substantiate the antiproliferative effects of EGAR, the apoptotic cells were monitored by annexin V adherence and caspase-3 activation. In viable cells, PS is located on the cytoplasmic surface of the cell membrane; in apoptotic cells, PS is translocated from the inner to the outer leaflet of the plasma membrane, thus exposing PS. Annexin V is a Ca2+–dependent phospholipids-binding protein with high affinity for PS. The binding of sophorodamine101-annexin V probe to PS that has translocated to the outer membrane cell produces red border around the cell under fluorescent microscope using red filter [Figure 1c]. Caspase-3 (CPP32) is a cytosolic protein that normally exists as a 32-kDa inactive precursor. It is cleaved proteolytically into a heterodimer when the cell undergoes apoptosis.[16] The cleavage of NucView™488 caspase-3 substrate by activated caspase-3 stains the cell nucleus green [Figure 1d]. The induction of apoptosis by EGAR was time dependent [Figure 2]. In MDA-MB-231 cells, an elevation in apoptosis-positive cells up to 26.5% was found after 3 hours and reached 48.3% after 6 hours of exposure to 20 μg/mL EGAR. In MCF-7, there were fewer apoptotic cells, EGAR at concentration of 30 μg/mL inducing apoptosis in 28.1% and 42.9% after 3 and 6 hours of treatment, respectively. The solvent controls did not increase the spontaneous apoptotic rate in the two malignant cell lines tested.

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