Detailed methods for in vitro/in vivo evaluation of anticancer drugs, with special reference to mistletoe extracts, have been reviewed. Mistletoe extracts have been shown to possess significant antitumor activity, in vivo, against murine tumors, Lewis lung carcinoma, colon adenocarcinoma 38 and C3H mammary adenocarcinoma 16/C. Methods for the extraction of biologically active alkaloids from mistletoe and their anticancer activities are presented. The possible origin of alkaloids in mistletoe plants, and their contributions towards a mechanism of anticancer activities of mistletoe axtracts, are proposed.