Dichloromethane fractions of Scrophularia oxysepala extract induce apoptosis in MCF-7 human breast cancer cells

Behnaz-Alsadat Hosseini; Ardalan Pasdaran; Tohid Kazemi; Dariush Shanehbandi; Hadi Karami; Mona Orangi; Behzad Baradaran

doi:10.17305/bjbms.2015.1.226

Authors

Behnaz-Alsadat Hosseini Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Ardalan Pasdaran
Tohid Kazemi
Dariush Shanehbandi
Hadi Karami
Mona Orangi
Behzad Baradaran

DOI:

https://doi.org/10.17305/bjbms.2015.1.226

Keywords:

Scrophularia oxysepala, cytotoxicity, MCF-7 cells, apoptosis, Real Time PCR

Abstract

Breast cancer is a prevalent malignancy among women, especially in developing countries. A large number of anticancer agents with herbal origins have been reported. Hence, herbals may play an essential role in prevention and treatment of cancers. We investigated cytotoxic effects of dichloromethane fractions of Scrophularia oxysepala extract on the MCF-7 breast cancer cell line. The cytotoxic activity of Scrophularia oxysepala fractions on the MCF-7 cells was assessed using Trypan Blue dye exclusion and MTT (3-(4, 5-dimetylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide) assays. In addition, apoptosis induction was determined using TUNEL (terminal deoxy transferase (TdT)-mediated dUTP nick- end labeling) assay and DNA fragmentation analysis. Quantitative Real-Time PCR was also used for analyzing the changes in Caspase-3, Caspase-9, and Bcl-2 genes’ expression. Results revealed an effective inhibition of growth and viability in MCF-7 cells treated with dichloromethane fractions. Cell death assay and DNA fragmentation analysis using the TUNEL test confirmed the induction of apoptosis in the MCF-7 cell line. Further, the fractions have resulted in an increased expression of Caspase-3and Caspase-9 mRNA, which highlights the possibility of apoptosis in the treatments. The expression study of Caspase-9 mRNA confirmed that, the fractions have triggered apoptosis via intrinsic mitochondrial pathway. In summary, fractions of Scrophularia oxysepala extract were found to be promising in growth inhibition and induction of apoptosis in MCF-7 breast cancer cells.

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