Theanticancer effects ofphytohormones ofcytokinin nature are similar to those ofmedicinal mushrooms, which are able to synthesize cytokinins in large amounts. To determine theantiproliferative effect ofcrude extracts and cytokinin fractions from themycelial biomass ofseven fungi species on colon cancer cells in vitro. Cytokinin content in mycelial biomass ofGanoderma lucidum, Lentinula edodes, Trametes versicolor, Pleurotus ostreatus, Morchella esculenta, Hericium coralloides, and Fomitopsis officinalis was determined by high performance liquid chromatography mass spectrometry. Theantiproliferative effect ofthemushroom extracts on thehuman colon adenocarcinoma Colo 205cells was assessed by MTT-test. Thecontent ofcytokinins (trans-zeatin, zeatin riboside, isopentenyladenosine, isopentenyladenine and zeatin-O-glucoside) was determined in themycelial biomass ofthemedicinal macromycetes. Zeatin-type hormones prevailed in all species, though trans-zeatin was themost abundant in H.coralloides and M.esculenta. In P.ostreatus, only zeatin-O-glucoside was detected. Thelowest IC50was found forboth thecytokinin fraction (0.21μg/ml) and thecrude extract (0.17μg/ml) from mycelial biomass ofH.coralloides. F.officinalis also demonstrated high antiproliferative effect against Colo205cells: IC50was 0.9μg/ml forthecrude extract and almost twice lower forthecytokinin fraction. In thestudied concentration range (0.016-2μg/ml), thecrude extracts from G.lucidum and M.esculenta and thecytokinin fraction from L.edodes did not reach IC50values. Thepresent study showed that crude extracts and/or cytokinin fractions ofseveral medicinal Basidiomycetes species are capable to inhibit proliferation ofcolon cancer cells in vitro. Crude extract cytotoxicity ofH.coralloides, P.ostreatus and T.versicolor was higher than that ofcytokinin fraction while antiproliferative effect ofcytokinin fraction from F.officinalis was higher than that in its crude extract.