Homo sapiens solute carrier family 35 member F2 (SLC35F2) is highly homologous to the lung squamous cell cancer-related gene, LSCC3, which is highly expressed in lung squamous cell tumour tissues. However, the clinical implication of the SLC35F2 gene in tumour development and progression remains unclear. An affinity-purified polyclonal antibody raised against the human SLC35F2 peptide was used in the immunohistochemical analysis of a non-small cell lung cancer (NSCLC) tissue microarray of human NSCLC (n=129). SLC35F2 protein was also analysed with the same antibody using Western blotting. Total RNAs were extracted from tumour tissues (n=43) and from laser-dissected tumour cells (n=9). SLC35F2 gene expression was detected by fluorescent real-time quantitative PCR and compared to the expression in the corresponding adjacent normal lung tissues. It was found that both the SLC35F2 protein (by IHC analysis) and the SLS35F2 gene transcripts (by Q-PCR analysis) were expressed at significantly higher levels in the NSCLC tumour tissues than in the corresponding adjacent normal lung tissues (p<0.001 and =0.015, respectively). There was a significant correlation between the SLC35F2 transcript and pathological staging (r=0.219, p=0.029), although the correlation between SLC35F2 protein and the staging was not significant (r=0.175). SLC35F2 was highly expressed in NSCLC tissues and the levels of expression, in particular the levels of the SLC35F2 transcript, were associated with NSCLC pathological staging. SLC35F2 appears to have a significant prognostic value in NSCLC.