Esophageal squamous cell carcinoma (ESCC) is a life‑threatening cancer with increasing incidence worldwide. MicroRNAs (miRs) have been reported to be involved in the progression of various types of cancer. In previous studies, the expression of miR‑378a‑3p was shown to be reduced in ESCC tissues. However, the mechanism underlying the effect of miR‑378a‑3p in ESCC remains to be elucidated. By employing a reverse transcription‑quantitative polymerase chain reaction, miR‑378a‑3p expression was tested in ESCC tissues and cell lines. In addition, the effects of miR‑378a‑3p on cell viability, proliferation, apoptosis, migration and invasion were studied using an MTT assay, an EdU assay, flow cytometry analysis, wound healing analysis and a Transwell assay. In the present study, the level of miR‑378a‑3p was significantly downregulated in ESCC clinical tissues and cell lines (EC109 and KYSE150). In addition, the overexpression of miR‑378a‑3p suppressed the viability, proliferation, migration and invasion of the ESCC cells. The upregulated expression of miR‑378a‑3p also increased the expression levels of B‑cell lymphoma 2‑associated X protein and caspase‑3, and decreased the expression levels of matrix metalloproteinase (MMP)‑2 and MMP‑9, which attenuated ESCC tumorigenesis. Furthermore, Rab10 was confirmed to be a direct target gene of miR‑378a‑3p, and was negatively affected by miR‑378a‑3p. The silencing of Rab10 revealed antitumor effects in ESCC cell lines, and the expression of miR‑378a‑3p was negatively correlated with that of Rab10 in ESCC. Collectively, miR‑378a‑3p may act as a tumor‑suppressor in ESCC cells through negatively regulating Rab10.