ANTIPROLIFERATIVE AND APOPTOTIC EFFECT OF METHANOLIC EXTRACT OF EDIBLE MUSHROOM Agaricus bisporus AGAINST HELA, MCF-7 AND MDA-MB-231 CELL LINES OF HUMAN CANCER AND CHEMOPROFILE BY GC-MS

The edible and wild mushrooms are recognized as important source of anticancer agents. Our aims are to harness methanolic extracts of Agaricus bisporus (ABME) and to test for anticancer effects in HeLa, MCF-7 and MDA-MB-231 cell lines. To test anticancer effect of this extract, we adopted some methodology such as cell morphology study by bright field microscope, nuclear morphology study by DAPI staining under inverted fluorescence microscopy, apoptosis assay by fluorescence technique, MTT assay, lactate dehydrogenase assays, and expression of apoptotic and anti-apoptotic genes by Western blotting. The results showed that under a phase contrast microscope, the cells from the three cancer cell lines exposed to 500–1000 μg/ml concentrations of ABME for 24 h, were round and shrunken. DAPI staining revealed that nuclei in the three treated cell lines were condensed, fragmented and deeply stained. MTT assay revealed that the growth of all three cell lines was inhibited at all tested concentrations of ABME; at 1000 µg/mL, the percentages of inhibition in HeLa, MCF-7 and MDA-MB-231 cell lines were > 80%. Under 500 µg/mL ABME exposure, LDH leakage in all three cell lines; increased expression of pro-apoptotic genes (Caspase 3, 9) and Bax; and decreased expression of BcL2 in MCF-7 cells validated the anticancer properties. The p53 gene was up-regulated by ABME. The percentage inhibition of migration of MCF-7 cells treated with ABME was 75.25±5.23. The total antioxidant content of ABME was 3.616718±0.24123 mg/ dry weight; the EC50 was 95.653±9.54067 µg/mL for DPPH, and the FRAP method indicated a free radical scavenging capacity of it. GC-MS analysis of the extract showed 102 compounds. In conclusion, A. bisporus thus contains anticancer compounds, and its consumption may prevent cervical and breast cancer.