Lung cancer remains the leading cause of cancer-associated mortality in China and worldwide. Increasing numbers of studies have demonstrated that microRNAs (miRNAs/miRs) have vital functions in numerous developmental processes and tumorigenesis. The aim of the present study was to investigate miR-154 expression in non-small cell lung cancer (NSCLC), and to explore the roles of miR-154 in the carcinogenesis and progression of this cancer. Reverse transcription-polymerase chain reaction (RT-qPCR) was performed to detect miR-154 expression in NSCLC tissues and cell lines. In addition, cell proliferation assay, migration and invasion assays were adopted to investigate the functional roles of miR-154 in NSCLC. Bioinformatics analysis, luciferase reporter assay, RT-qPCR and western blot analysis were used to explore the potential targets of miR-154 in NSCLC. According to the results, miR-154 was significantly downregulated in NSCLC tissues and cell lines. Restoration of miR-154 expression inhibited proliferation, migration and invasion of NSCLC cells. In addition, B-cell-specific Moloney murine leukemia virus insertion site 1 (BMI-1) was identified as a direct target gene of miR-154 in NSCLC. In conclusion, miR-154 may function as a tumor suppressor in NSCLC, partly by regulating BMI-1, and the modulation of miR-154 expression represents a potential strategy for the treatment of NSCLC patients.