Anticancer potential of methanol extract of seeds of Artocarpus hirsutus in human breast cancer cell lines

Abstract

Artocarpus hirsutus, belonging to the family Moraceae, is used traditionally for the treatment of skin problems, diarrhoea, snake bite and ulcers. In the present study, the methanol extracts of fruit seeds of A. hirsutus (MAH) (Wild jack) were investigated for the presence of phytochemicals and anticancer activities against human breast cancer cell lines in vitro. Methanolic extraction was done using soxhlet. The phytochemicals were determined by conventional qualitative tests and GC-HRMS. Cytotoxicity study was conducted using MTT assay. Apoptotic changes produced by the plant extract in the treated cells were determined by acridine orange ethidium bromide and hoechst staining. Apoptotic pathway was analysed using JC-1 staining and molecular expressions of Bcl-2. Protein and gene expressions of Bcl-2 were studied by performing western blot and RT-qPCR respectively. In the cytotoxicity study, MAH exhibited significant (p<0.05) reduction in per cent cell viability. The IC₅₀ value was found to be 65.47±9.51 μg/mL in MDA-MB-231 cells and 64.05±9.3 μg/mL in MCF-7 cells. Microscopic studies in both the cells after MAH treatment revealed morphological and nuclear alterations which were characteristics of intrinsic pathway of apoptosis. Significant (p<0.05) down regulation in the expression of anti-apoptotic gene and protein Bcl-2 was observed which confirmed the intrinsic pathway of apoptosis. The present study revealed a concentration dependent anticancer activity of methanol extract of seeds of A. hirsutus mediated through intrinsic pathway of apoptosis.