Background: Esophageal cancer (EC) is a common malignancy that claims millions of lives globally each year. Impairment of esophageal carcinoma cell proliferation could provide a therapeutic approach to treating EC. MiR-449a-5p has been put forward as a crucial tumor suppressor in multiple cancers. In this study, we aimed to determine the mechanism underlying the regulating effect of miR-449a-5p on EC cell proliferation. Methods: The level of miR-449a-5p in esophageal squamous cell carcinoma (ESCC) tissues and adjacent normal esophageal squamous tissues was detected using real-time polymerase chain reaction. The protein level of B-cell lymphoma 2 (Bcl-2) was measured by western blot. MTT and colony formation assays were carried out to assess Eca-109 cell proliferation. Results: The level of miR-449a-5p in ESCC tissues was reduced in comparison with that in adjacent normal tissues. Down-regulation of miR-449a-5p promoted the proliferation of ESCC cells. Bcl-2 was established as a target gene of miR-449a-5p, and its silencing resulted in the reversal of the effects of miR-449a-5p inhibitor on the proliferation of Eca-109 cells. Conclusion: Overexpression of miR-449a-5p inhibited the proliferation of Eca- 109 cells via negative regulation of Bcl-2.