In silico identification of natural Anticancer product and their efficacy in Breast cancer cells and cancer stem like Cells

Abstract

Breast cancer is the most commonly diagnosed lethal cancer in women worldwide. newlineNotch signaling pathway is directly linked to breast cancer recurrence and newlineaggressiveness. Natural remedies are becoming a prime choice to overcome against newlinecancer due to lesser side effect and cost-effectiveness. Literature survey and in silico newlinestudy identified Bulbine frutescens (Asphodelaceae), Kurarinone (KU) and 3-O-(E)-p- newlinecoumaroylbetulinic acid (CB) as lead plant product/phytochemicals. Methanolic and newlinehexane extract of B. frutescens (BME and BHE respectively), KU and CB were studied newlinefor their anticancer activity and notch signaling pathway inhibitory potential in breast newlinecancer cells. Moreover, KU and CB were also studied for their effect in mammosphere. newlineLiterature-based identification of methanol soluble phytochemicals of B. frutescens and newlinein silico docking study revealed Bulbineloneside D as a potent notch signaling inhibitor newline(and#978;-secretase). In silico docking potential of KU and CB were equal to standard gamma newlinesecretase inhibitor DAPT (-8.74 kcal/mol). KU-gamma secretase complex showed newlinelower RMSD value, marginal fluctuation in Radius of gyration (Rg), more number of newlineinter hydrogen bonding, and stable secondary structure of the protein which indicates newlineKU as candidate gamma secretase inhibitor (GSI). B. frutescens extracts (IC50 4.8 newline28.4 and#956;g/ml), Kurarinone (IC50 0.43-3.42 µM) and CB (IC50 0.99-5.88 µM) significantly newlinedecreased cell viability in MDA-MB-231 and T47D cells in time dependent manner. B. newlinefrutescens, KU and CB induced cell cycle arrest at G1 phase in MDA-MB-231 and T47D newlinecells. RT-PCR analysis of cell cycle (cyclin D1, CDK4, and p21) and apoptosis newlinemodulating genes (caspase 3, Bcl2 and survivin) revealed upexpression of p21, and newlinecaspase 3, and down expression of cyclin D1, CDK4, Bcl2 and survivin genes in test newlineextract/phytochemicals treated breast cancer cells. Western Blot analysis showed newlinereduced expression of cyclin D1 and increased procaspase 3 protein expression in newlineextract/phytochemicals treated breast cancer cells in t