To investigate a possibility of producing the antitumor component by shake culture method, the mycelia of Flammulina velutipes were cultured in flasks on a shaker at $26{sim}28^{circ}C$ at 180 rpm for seven days. The extract of the mycelia was concentrated under vacuum. The precipitate obtained by adding a three-fold volume of ethanol was centrifugated and freeze-dried after dialysis. The fraction was tested against sarcoma 180 in the mice. The inhibition ratio of the fraction against the tumor was 68.0% at the dose of 20 mg/kg/day for the period of ten days and the tumors in three of the mice were completely regressed. The results showed, therefore, that the antitumor component was produced by the shake culture method.