Golgi phosphoprotein 3 (GOLPH3) has been reported to be involved in the development of several human tumours. However, little is known about GOLPH3 expression and its clinical significance in non-small cell lung cancer (NSCLC). The present study was conducted to investigate the expression of GOLPH3 and its prognostic significance in NSCLC. Immunohistochemical analysis was used to determine the expression of GOLPH3 in 145 cases NSCLC tissue and their corresponding adjacent normal tissues. The results are the following: (1) The GOLPH3 protein was mainly located in the cytoplasm of NSCLC tissue; (2) GOLPH3 was highly expressed in 71.7% of NSCLC tissues versus 22.8% of adjacent tissues (P < 0.01); (3) GOLPH3 expression was significantly associated with tumour-node-metastasis (TNM) stage (P = 0.001), lymph node status (P = 0.014), and degree of differentiation (P = 0.018); (4) The Kaplan-Meier curve indicated that the patients with high GOLPH3 expression had significantly shorter survival than those with low GOLPH3 expression; and (5) Cox regression analysis demonstrated that the expression of GOLPH3, lymph node status, and TNM stage were independent prognostic predictors for NSCLC patients. High GOLPH3 expression is predictive of poor prognosis of NSCLC, implying that GOLPH3 may be a promising new target for targeted therapies for NSCLC.