Cervical cancer is the fourth most common female malignancy worldwide and represents serious health problem, especially in less developed countries. Metastasis is an important issue in cancer therapy. Very often alternative medical approach, such as fungotherapy, has been applied on many cancer patients. Mushrooms have been found to produce a wide range of complex metabolites, some of which have pivotal biological activity. An edible and medicinal mushroom Laetiporus sulphureus (Bull.) Murrill is known to be prolific producer of phenolic compounds, polyketides, terpenes and steroids, with potential medicinal applications in the treatment of various diseases such as cancer. This study aimed to analyze effects of L. sulphureus on cervical cancer cell line (HeLa). Cells were seeded for nitroblue tetrazolium test (NBT) and Wound healing tests, and treated with ethanolic mushroom extract in two selected concentrations (10 and 50 µg/mL). Effects were tracked after 24 h and 72 h for NBT assay, while for Wound healing test effects were evaluated 24 h of treatment. Extract induced acute prooxidative effect on HeLa cells, while the level of superoxide anion radicals fell below the control values after a longer time of exposure to the treatment. Ethanol extract of L. sulphureus had an antimigratory potential 24 h after treatment when compared to untreated cells, with a higher concentration (50 µg/ml) having a slightly stronger antimigratory activity. Laetiporus sulphureus is a very important source of significant biological potential and should be reconsidered for development of promising anticancer therapies.