Together, these results indicate that MT3‐MMP negatively regulates proliferation and aggressiveness of ESCC cells by blocking progression of cell cycle from G1 to S and then G2/M phase (e.g., through the restriction point of cell cycle), likely in association with up‐regulation of the endogenous Cdk inhibitors p21Cip1 and p27Kip1. Here, CDKN1B is linked to esophageal squamous cell carcinoma.