The invention discloses a traditional Chinese medicine composition with an antineoplastic effect, and a preparation method and an application thereof. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 15-30 parts of barbed skullcap herb, 15-30 parts of oldenlandia diffusa, 15-30 parts of rabdosia rubescens, 3-15 parts of radix sophorae flavescentis, 5-15 parts of curcuma zedoary, 5-15 parts of stir-baked fructus arctii, 5-15 parts of red ginseng, 5-30 parts of astragalus mongholicus, 6-12 parts of bighead atractylodes rhizome, 10-15 parts of poria cocos, 6-15 parts of radix paeoniae alba, 5-15 parts of Chinese angelica, and 3-20 parts of radix glycyrrhizae preparata. According to the traditional Chinese medicine composition, traditional Chinese medicine proportioning prescription is conducted by a theory of treatment based on syndrome differentiation according to a. traditional Chinese medicine theory; and clinical experimental results indicate that the traditional Chinese medicine composition has a better killing effect on cells of various tumors such as a gastric cancer, a liver cancer and a lung cancer, is wide in application scope, has no adverse effect clinically and is safe and reliable to take, and a new-generation antineoplastic drug is expected to be developed.