Objective:To investigate the anti-tumor effect and mechanism of CⅡ-3 extractive from Periplaneta americana on S180 sarcoma-bearing mice. Methods: S180 sarcoma-bearing mice were randomly divided into model group, positive control (CTX) group, high-dose, medium-dose and low-dose CⅡ-3 groups, and a normal control group, with 10 mice in each group. The pathological changes of tumor tissue in each group were observed after administration, and the tumor inhibition rate was calculated. ELISA kits were used to test the levels of vascular endothelial growth factor(VEGF) and telomerase(TE) in serum. The expression of p53, B-cell lymphoma 2(Bcl-2), cysteinyl asparate specific protease-3(Caspase-3), microvessel density (MVD) and cyclooxygenase-2(COX-2) protein in tumor tissue were detected by immunohistochemistry. Results: The tumor inhibition rates of CTX group, high-dose, medium-dose, and low-dose CⅡ-3 groups were 85.71%, 55.71%, 44.28%, 34.28%, respectively. The necrotic area of tumor tissue was obvious in high-dose and medium-dose CⅡ-3 groups. Compared with the model group, the TE activity was decreased in high-dose CⅡ-3 group(P<0.05), and the expression of p53, Bcl-2, MVD and COX-2 in tumor tissues of high-dose and medium-dose CⅡ-3 groups were significantly decreased(P<0.01), while the Caspase-3 expression was significantly increased(P<0.01). Conclusion: CⅡ-3 can significantly inhibit the growth of S180 sarcoma, and its mechanism may be related to the reduction of TE activity, the decrease of p53, Bcl-2, MVD, COX-2 expression, and the increase of Caspase-3 expression in tumor tissue.