The fungal strains were obtained by single colony isolation technique from the soil of the world’s unique proved independent selenium deposit. The anti-tumor activity was assayed by MTT method using MDA-MB-231cells, Hela cells and SMMC-7721 cells, respectively, together with morphological observation of these cells under inversed light microscope. The 73 terrestrial fungal strains were isolated from the soil samples collected from 6 different areas in Enshi area of HuBei Province, China. Among them, 32 strains produced fermentation products obviously inhibiting the Hela cells, 13 strains inhibiting the MDA-MB-231 cells and 9 strains inhibiting the SMMC-7721 cells with IC₅₀ value less than 90 µg/ml. These antitumor-metabolite-producing strains have provided fungal strains for further studies on the bioactive metabolites.