Objective To evaluate the sensitivity to 5 clinically commonly used anticancer drugs in vivo using the zebrafish xenotransplantation models of human lung cancer, stomach cancer, and liver cancer cells, respectively. Methods Zebrafish xenotransplantation models of A549 lung cancer cells, SGC-7901 stomach cancer cells and HepG2 liver cancer cells were established. The xenograft models of A549 cells were treated with three different doses of cis-platinum, paclitaxel, vinorelbine, endostar and bevacizumab, respectively. The SGC-7901 model was treated with three concentrations or doses of paclitaxel, irinotecan, hydroxyurea, cis-platinum and 5-fluorouracil, respectively. And the HepG2 model was treated with three concentrations or doses of adriamycin, gemcitabine, hydroxyurea, cis-platinum and 5-fluorouracil. The tumors were analyzed and quantified in vivo by fluorescence microscopy, and the inhibition rates of tumor growth with each drug were calculated and compared with the model control group for statistical significance. Results All of the tested anticancer drugs showed inhibitory effect on tumor cells in the zebrafish xenograft models with statistical significance in a dose-dependent manner. During the drug sensitivity test, the inhibition rate of bevacizumab on A549 lung cancer cells decreased in the order (65%) > cis-platinum (55%) > vinorelbine (40%) > endostar (39%) > paclitaxel (27%). As for the SGC-7901 stomach cancer cells, the tumor growth inhibition rate decreased in the order hydroxyurea (46%) > 5-FU (31%)=irinotecan (31%) > paclitaxel (26%) > cis-platinum (24%). And the therapeutic effect of cis-platinum on the HepG2 liver cancer cells decreased in the order (64%) > hydroxyurea (56%) > gemcitabine (46%) > adriamycin (45%) > 5-FU (38%). Conclusions Zebrafish xenotransplantation models of cancer cells are suitable for in vivo sensitivity test of anticancer drugs.