Aim To explore injury action and phenotype shift on mouse vascular smooth muscle cells by cigarette smoke extract(CSE）. Methods CSE was prepared, 2.5%, 5%, 10% and 5% of CSE was respectively used to treat mouse vascular smooth muscle cells(MOVAS cells）, and normal MOVAS cells were used as control group. Cell survival rate was detected by MTT assay. Intracellular reactive oxidative species(ROS)level was estimated by ROS kit. MOVAS cell cycle and apoptotic rate were respectively analyzed by flow cytometry instrument combining PI simple staining and Annexin V-PI double staining. SMemb and SMA mRNA expression of MOVAS cells were analyzed by real-time quantitative PCR, SMemb and SMA protein expression of MOVAS cells were measured by Western blot. Results After 2.5%, 5%, 10% and 20% CSE processed, survival rate of MOVAS cells was respectively 89.6%±8.6%, 65.2%±10.0%, 53.6%±6.9% and 43.1%±5.9%, the difference was statistically significant compared with the control group(P<0.05）, meanwhile, ROS level increased significantly. After 10% CSE processed, the percent of cells in G1 cycle was increased obviously, the difference was statistically significant(P<0.05）. Cell apoptosis rate also increased significantly, the difference was statistically significant(P<0.05）. The expression of SMemb as MOVAS synthetic symbol was significantly increased, while the expression of SMA as contractile symbol was reduced remarkably. Conclusion CSE can injure mouse vascular smooth muscle cells, and promote the transformation of MOVAS cells’ phenotype from synthetic to contractile.