Objectives: The aerobic glycolysis is a characteristic of cancer. α-enolase (ENO1) is an important glycolytic enzyme and is suggested to play a role in the development of tumor. The purpose of this study was to investigate the effects of ENO1 on growth, invasion, and metastasis of human cervical cancer cells. Methods: We knocked down gene coding ENO1 in Hela and SiHa cells with lentivirus, which was confirmed by western-blot. Following downregulation of ENO1, the drug sensitivity, invasion, metastasis, tumourigenic abilities of ENO1-altered cells were investigated. Results: Our studies demonstrated that downregulation of ENO1 enhanced the sensitivity of Hela and SiHa cells to paclitaxel and cisplatin and decreased their tumorigenicity. In addition, the abilities of invasion, metastasis, and tumourigenesis were decreased in ENO1-silenced Hela and SiHa cells. Conclusions: ENO1 is involved in the tumorigenicity, invasion, metastasis, and drug sensitivity of Hela and SiHa cells.