Antitumor activity of a polysaccharide fraction (GF-1) extracted from cultured fruiting bodies of a fungus, Grifola frondosa, was examined on allogeneic and syngeneic tumors in mice. GF-1 had a marked inhibitory activity against the growth of subcutaneously (s.c.) inoculated Sarcoma 180 by the intraperitoneally (i.p.) injection at 0.5-5.0 mg/mouse for 10 successive days. A significant antitumor activity was also observed when GF-1 at 4.0 mg/mouse was i.p. injected successively on days +1-+5, +7-+11, +14-+18 or +21-+25 if the tumor cells were inoculated s.c. on day 0. Similar results were obtained by a single i.p. injection of GF-1 at 2.0 mg/mouse on day +1, +7, +14 or +21. When GF-1 was injected i.p., intravenously (i.v.) or intratumorally (i.t.), all of them showed an equivalent level of higher inhibitory activity (inhibition ratio; over 90%). However, the oral (p.o.) administration was not effective. The pretreatment of mice with GF-1 at 2.0 or 4.0 mg/mouse for 5 times before tumor inoculation did not show a significant antitumor activity. In addition, GF-1 administered i.p. at varying times before and/or after the tumor inoculation showed no antitumor activity against ascites form of Sarcoma 180. In the syngeneic systems, GF-1 exhibited an antitumor activity against solid form of Meth A fibrosarcoma in BALB/c mice and MM46 carcinoma in C3H/He mice.