Abstract
Objective: To evaluate the leaf extract of Abrus precatorius as a potential therapy for breast cancer treatment.Methods: Aqueous leaf extracts of A. precatorius was prepared by the process of maceration. The collected filtrate was further partitioned successively into five solvent fractions starting from water, hexane, chloroform, ethyl acetate and butanol by solvent fractionation method using separating funnel. Finally, all the five fractions were subjected to cytotoxic activity assay. The molecular mechanism underlying cytotoxicity was determined by using various approaches viz., 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Fluorescence-activated cell sorting (FACS) analysis, Western blot analysis, casp-glow assay and Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis.Results: Among five fractions only ethyl acetate fraction of A. precatorius (EAF-AP) showed significant cytotoxic activity on MDA-MB-231 cells, with an IC50 value of 47.3 µg/ml. Apoptosis was confirmed by the appearance of Sub G0/G1 (apoptotic) peak by FACS analysis. Western blotting results clearly indicated cleavage of Caspase-3 and PARP. Casp-glow assay confirmed activation of caspase-3, an important mediator of apoptosis. Semi-quantitative RT-PCR analysis showed an up-regulation of pro-apoptotic genes (p21, p53 and Bax) and down-regulation of the anti-apoptotic Bcl-2 gene, which is an important hallmark of an apoptosis.Conclusion: All these results indicate that EAF-AP, induced apoptosis in MDA-MB-231 cells, making A. precatorius a potentially good candidate for anticancer drug development.
Highlights
Cancer is a multifaceted devastating disease that arises as a result of a multistep process called carcinogenesis affecting millions of people per year and poses both economic and psychological challenges [13]
Treatment of MDA-MB-231 cells for 48 h with an EAF-AP showed a very significant and higher cytotoxic activity with an IC50 value of 47.3 μg/ml. These results indicated that EAF-AP has a potent cytotoxic activity on MDA-MB-231 breast cancer cells
Microscopic examination of MDA-MB-231 cells treated with EAF-AP showed significant morphological changes such as shrinkage and detachment from the surface
Summary
Cancer is a multifaceted devastating disease that arises as a result of a multistep process called carcinogenesis affecting millions of people per year and poses both economic and psychological challenges [13]. It is a major public health problem that continues to be a second leading cause of death in humans all over the world. It has been reported around 14.1 million people were detected with cancer and 8.2 million people died from cancer [4]. Worldwide breast cancer mortality among females was 23 % (1.38 million) of the total new cancer cases and 14% (458,400) of the total cancer deceases recorded in 2008 [7]
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