Introduction: Cancer is a non-communicable disease with over 18.1 million new cases and 9.6 million deaths annually according to WHO. Breast cancer is the number two highest count type of cancer trailing behind lung cancer. Treating cancer is expensive and have various side effects. Active components found in plants or fungi that have antioxidant and cytotoxic activity towards cancer cells, could be an alternative for anticancer. One of the fungi that is potentially developed as an anticancer, are the genus of Auricularia sp. also known as black-white fungus. This study aims to determine the phytochemicals components, antioxidant activity and cytotoxic effect of the Auricularia sp. towards MCF-7 breast cancer cells. Methods: Dried black-white fungus of Auricularia sp. grinded into a fine powder. Then, multilevel maceration is done with the n-hexane, ethyl acetate, ethanol as solvents. The extracts of black-white fungus undergo phytochemical screening and thin layer chromatography (TLC), followed by measuring antioxidant and evaluating the cytotoxic activity towards MCF-7 breast cancer cells. Results: black-white fungus of Auricularia sp. contained secondary metabolites of flavonoids, alkaloids, and triterpenoids and a total of 17 other phytochemical components. Ethyl acetate extract of black-white fungus showed a weak antioxidant activity towards DPPH free radical with IC50 of 215.51 μg/mL and a very active cytotoxic activity on MCF-7 cells with IC50 of 0.21 μg/mL. On the other hand, ethanol and n-hexane extracts of black-white fungus are categorized with an active cytotoxic activity on MCF-7 cells with IC50 of 29.28 μg/mL and 50.39 μg/mL, respectively. Conclusion: Black-white fungus Auricularia sp. that had anticancer activity towards breast MCF-7 cells should be considered as an alternative treatment for breast cancer thera © 2022 Phcogj.Com. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.