Lung cancer is the leading cause of cancer-related death worldwide, and current therapeutic strategies such as chemo- and radiotherapy remain unsatisfactory due to drug resistance or adverse reactions. Therefore, exploring the anticancer effects of natural components is getting really important. In this study, we performed antioxidant tests on strawberry methanolic extract and determined its total polyphenol and flavonoid content. The MTT assay was used to evaluate the toxicity of strawberry extract from 1.5 to 15.0 mg/mL on A549 cells at different times. ROS, apoptosis and cell cycle analysis were also evaluated in strawberry-treated cells. The results showed that the cytotoxicity of strawberry extract on A549 was time- and dose-dependent. Furthermore, strawberry extract significantly increased the ROS and apoptosis levels of A549 cell and induced Sub G1 cell cycle arrest after 48 hours of treatment. The findings suggest that strawberry extract has great potential to inhibit lung cancer.