Abstract: ob＜x＞jective: To investigate the antitumor effect of LIM and SH3 protein 1 gene (LASP1) interfering adenovirus (Ad-LASP1-shRNA) on non-small cell lung cancer. Methods: Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect LASP1 ex＜x＞pression levels in 56 NSCLC tissues; Ad-LASP1-shRNA infected A549 cells for 48 h; The ex＜x＞pression level of mRNA and proteins in transfected A549 cells were analyzed by qRT-PCR and Western blot; Scratch test and Transwell method to detect the migration and invasion ability of A549 after transfection; Build a A549 transplanted model, the tumor volume was measured regularly after injected Ad-LASP1-shRNA and PBS. Results:The relative ex＜x＞pression of LASP1 in cancer tissue and adjacent tissue was 1.58±1.02 and 0.84±0.72 (P<0.001). qRT-PCR and Western blot suggested that Ad-LASP1-shRNA can effectively inhibit the ex＜x＞pression of LASP1. The migration and invasion ability of A549 cells infected Ad-LASP1-shRNA decreased significantly after 24 h, and the number of invasive and migratory cells was significantly lower than that of the control group (P<0.05); Ad-LASP1-shRNA can significantly inhibit the growth of transplanted tumors. The tumor volume of Ad-LASP1-shRNA group was (334.5±32.6) mm3, significantly smaller than that of PBS group (833.9±61.3) after 5 weeks of inoculation(P<0.001).