Aim: To evaluate the anti tumor effect of compound DAT-230, a novel analog of Combretastatin A-4, and explore the mechanism, in vitro.Methods: Cytotoxicity was measured by MTT method.The Cellular microtubule network was visualized by immunofluorescence and Western blotting.Cell cycle distribution was measured using PI staining by flow cytometry.The mitochondria membrane potential was detected by Rho123 staining and flow cytometry.The early stage of apoptosis was monitored by Annexin V-fiuorescein and PI double staining.MDC staining was used to detect the autophagy ratio.All the proteins expression and activation were measured by Western blotting in HT-1080 and SGC-7901 cells.Results: A novel synthesized compound DAT-230, inhibited the proliferation of caner cells including KB (IC50 =0.05 μM), A549 (IC50 =0.5 μM), H460 (IC50 =0.25 μM), SGC-7901 (IC50 =0.08 μM), HT1080 (IC50 =0.03 μM) and HEpG-2 (iC50 =0.1 μM).Tubulin polymerization assay, Western blot and immuno fiuorescence experiments suggested that DAT-230 inhibit microtubule assembly at cellular levels similar to the effect of combretastatin-A4.DAT-230 induced time-and dose-dependent G2/M cell cycle arrest, and finally resulted in apoptosis in KB, A549, SGC-7901, HT-1080 and H460 cells.Further, in HT-1080 and SGC-7901 cells, accompanying with G2/M cell cycle arrest, cyclin B1, cdc25c and phosphorylation of Cdc2 at Thrl4 and Tyr15 were up regulated, while Cdc2 itself and CDK7 werent regulated.The protein level alteration and the abnormal mitosis spindle were observed, denoting the M phase arrest in DAT-230-treated cells.Moreover, DAT-230 also induced apoptosis of HT1080 cells via mitochondria apoptotic pathways, as indicated by a decrease in mitochondrial membrane potential (MMP), plasma membrane translocation of phosphatidylserine, cleavage of poly (ADP-ribose) polymerase, procaspase-3 and pro-caspase-9, and changes of the expression ratio of Bcl-2/Bax.Taken together, all the data demonstrated that DAT-230 exhibited its anti tumor activity through disrupting the microtubule assembly, causing cell cycle arrest, and consequently inducing apoptosis in HT-1080cells.Conclusions: The novel compound DAT-230 is a promising microtubule inhibitor that has great potentials for therapeutic treatment of various malignancies.These data suggest that DAT-230 produces anti-tumor effect via induction of G2/M cell cycle arrest and apoptosis.