Objective To detect the expression of survivin in normal lung and non-small cell lung cancer (NSCLC) tissues, and investigate the relationship of its expression with the clinical pathological characteristics of NSCLC as well as the cancer cell cycle. Methods Immunohistochemical SP staining was used to detect the expression of survivin in 43 samples of NSCLC tissues and 10 samples of normal lung tissues. The relationship of survivin expression and clinicopathological characteristics of NSCLC was analyzed. In order to evaluate the correlation of survivin expression with cancer cell cycle, flow cytometry was used to detect the cells isolated from 6 NSCLC samples with 6 samples without survivin expression to calculate the values of S-phase fraction (SPF) and proliferation index (PI). Results (1) The positivity rate of survivin expression was 51.16% in NSCLC tissues, and 0.00% in normal lung tissues, with significant difference between them (P<0.05). (2) The expression of survivin was obviously correlated with patient’s gender and lymph node metastasis (P<0.05), but not with patient’s age, pathological classification and T staging (P>0.05). (3) The results of flow cytometry indicated that there were statistical significances in PI value (11.80±4.25 vs 5.68±2.56) and SPF value (13.93±3.21 vs 6.44±2.52) between the cells isolated from the NSCLC samples with or without survivin expression (P<0.05). Conclusion (1) The expression of survivin is significantly higher in the NSCLC tissue than in normal lung tissue. (2) Survivin expression is obviously correlated with gender and lymph node metastasis. (3) The PI and SPF values are significantly higher in survivin positive than negative NSCLC cells, which indicate that survivin positive expression promotes the proliferative activity of tumor cells through inhibition of apoptosis.