AbstractLong non‐coding RNAs (lncRNAs) have been shown to be dysregulated in a variety of malignant and non‐malignant lesions including non‐functioning pituitary adenomas (NFPAs). In the current experimental study, we have selected six lncRNAs, namely MAPKAPK5‐AS1, NUTM2B‐AS1, ST7‐AS1, LIFR‐AS1, PXN‐AS1 and URB1‐AS1 to assess their expression in a cohort of Iranian patients with NFPA. MAPKAPK5‐AS1, PXN‐AS1 and URB1‐AS1 were shown to be over‐expressed in NFPA tissues compared with control samples (Expression ratios (95% CI) = 10 (3.94–25.36), 11.22 (4.3–28.8) and 9.33 (4.12–21.12);pvalues < 0.0001, respectively). The depicted ROC curves showed the AUC values of 0.73, 0.80 and 0.73 for MAPKAPK5‐AS1, PXN‐AS1 and URB1‐AS1, respectively. Relative expression level of PXN‐AS1 was associated with tumour subtype (pvalue = 0.49). Besides, relative expression levels of MAPKAPK5‐AS1 and LIFR‐AS1 were associated with gender of patients (pvalues = 0.043 and 0.01, respectively). Cumulatively, the current study indicates the possible role of MAPKAPK5‐AS1, PXN‐AS1 and URB1‐AS1 lncRNAs in the pathogenesis of NFPAs.