Abstract: OBJECTIVE To optimize extraction process of sporoderm-broken spores of Ganoderma lucidum triterpenoids and evaluate anti-cancer effect of total triterpenoids from spoderm-broken spores of Ganoderma lucidum in vitro. METHODS Orthogonal test was used to optimize the extraction process of triterpenoids and cell viability was detected by MTT assay. Besides, the apoptosis rate of cells was detected by flow cytometry. RESULTS The optimum extraction conditions were as follows:ethanol concentration:95%, the ratio of liquid to material ratio:60:1, temperature:85℃, time:2 h, frequency:2. According to this condition, the content of triterpenoids of Ganoderma lucidum was up to 6.45%. The IC₅₀ values of triterpenoids of Ganoderma lucidum on human colon cancer HCT116 cells, human lung cell line A549 cells and human breast cancer MDA-MB-231 cells were 1.29, 1.68, 3.81 mg·mL^-1, respectively. Compared with the control group, apoptosis of HCT116 cells treated with a various concentration of total triterpenoids of Ganoderma lucidum (0.64, 1.6, 4.0, 10 mg·mL^-1) at 36 h was significantly induced. CONCLUSION The extraction process is feasible, which can improve extraction efficiency. The MTT data indicates that triterpenoids of Ganoderma lucidum has anti-proliferation activity of human colorectal cancer, human breast cancer and huaman lung cancer cells in vitro, and the results show that triterpenoids of Ganoderma lucidum can significantly induce apoptosis of HCT116 cells.