The abnormal expression of microRNA-221 was detected in several cancers and some studies had indicated that microRNA-221 was associated with cancer prognosis. This study was aimed to evaluate the prognostic significance of microRNA-221 in non-small cell lung cancer (NSCLC). Quantitative real-time polymerase chain reaction (qRT-PCR) was used for detecting the relative expression levels of microRNA-221 in the pathological tissues and corresponding normal tissues of 104 NSCLC patients. The relationship between the expression levels and the clinical features was estimated by Chi-square method and the overall survival of patients at different expression levels was demonstrated by Kaplan-Meier method. Cox regression analysis was used to evaluate the prognostic significance of microRNA-221. The relative expression levels of microRNA-221 in the pathological tissues were remarkably higher than that in the corresponding normal tissues (1.71 vs 1.07, P = 0.000). The expression level was associated with lymph node metastasis (P = 0.001). The results of Kaplan-Meier method indicated that patients with high expression level of microRNA-221 had shorter overall survival time than those with low expression level (36.8 vs 45.2 months, P = 0.001). Moreover, Cox regression analysis suggested that microRNA-221 was a useful independent biomarker for NSCLC prognosis (HR = 1.873, 95 % CI = 1.267-2.768, P = 0.002). The aberrant expression of microRNA-221 is associated with NSCLC progression and it might be a potential biomarker for NSCLC prognosis.