Abstract: Objective To explore the inhibitory effect of bafilomycin A1（Baf-A1）on cell proliferation and autophagy of touge squamous cell carcinoma Tca8113 cells. Methods Tca8113 cells were cultured with Baf-A1（0.1，0.5 μmol／L）, and cells without treatment were set as control group. The cell proliferation was determined at 0，24，48 and 72 h by MTT assay in control group and Baf-A1 group. Flow cytometry was applied to measure cell cycle treated with 0.5 μmol／L Baf-A1 for 72 h. Western blotting and immunofluorescence were used to analyze autophagy marker LC3B protein expression. Autophagosome formation was observed by transmission electron microscope. Results Baf-A1 could dose-dependently inhibit the proliferation of Tca8113 cells. The highest proliferative inhibition was observed at 72 h after cells treated with 0.5 μmol/L BafA1. Compared with control group, Tca8113 cells in Baf-A1 group increased in sub-G1 and G1 phase（P＜0.05）, and decreased in S and G2/M phase（P＜0.05）. LC3B-II expression of Baf-A1 group was 1.83±0.23，which was significantly higher than 0.79±0.18 of control group（P＜0.05）. Immunofluorescence showed that little LC3B could be seen in control group, while large amount of LC3B puncta was observed in Baf-A1 group after 72 h. Compared with control group，large amount of autophagosomes were observed and the formation of autolysosomes increased in Baf-A1 group. Conclusion Baf-A1 has a toxic effect on the proliferation of Tca8113 cells. It may be related to cell cycle arrest and autophagy inhibition.