There are few studies on the role of ring finger protein (RNF)180 in non-small cell lung cancer (NSCLC). The present study investigated the expression of RNF180 in NSCLC and its associations with the clinical factors and prognosis of NSCLC. The mRNA and protein expression levels of RNF180 were detected via reverse transcription-quantitative PCR and western blotting. Methylation-specific PCR (MSP) analysis was utilized to detect the methylation of RNF180. RNF180 expression levels were analyzed via immunohistochemistry. The protein and mRNA expression levels of RNF180 were lower in NSCLC cell lines compared with in the non-tumor cell line. Immunohistochemistry revealed that 64 patients that were negative for RNF180, while MSP detection analysis demonstrated that 60 patients exhibited RNF180 promoter methylation. The methylation status of RNF180 was significantly associated with RNF180 expression level. Among all factors evaluated, logistic regression analysis indicated that only T stage was significantly associated with RNF180 expression. Cox multivariate analysis demonstrated that RNF180 expression was an independent predictor of overall survival in patients with NSCLC. Methylation in the promoter of RNF180 was shown to reduce its expression levels. In summary, low RNF180 expression levels were associated with poor biological behaviors, thus RNF180 expression level may be used as a clinical marker to predict the prognosis of patients with NSCLC.