Four- to five- week-male nude mice were purchased from Gem Pharmatech Co., Ltd and housed on a 12 h light–dark cycle under pathogen-free conditions. The animal experiment was approved by the Committee on the Ethics of Animal Experiments of the Yantai Institute of Materia Medica, and was performed in strict accordance with the guidelines of the National Research Council's Guide for the Care and Use of Laboratory Animals. HCT-116 cells were harvested, 2 × 106 (link) cells per 200 μL were injected on the flank of the mice. Tumor growth was measured, when the average tumor volume reached to 200 mm3, the mice were randomized into six groups (n = 5). The mice in the treatment groups were treated with DM1 (1 mg kg−1), LWJ-M30 (1 mg DM1 equiv. kg−1) twice a week in the tail. The mice in control group were administrated with 5% glucose solution. Tumor size was measured three times a week using a caliper and the volume was calculated as
After 21 days treatment, the mice were euthanized and the tumors were excised and weighed.