Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer. LncRNA HOTAIR (HOx Transcript AntIsense RNA) and Galectin-3 are involved in PTC. This study explored the clinical effect of lncRNA HOTAIR/Galectin-3 on PTC patients.Subjects were assigned into PTC (160 cases) and benign thyroid tumor groups (150 cases). Fasting peripheral venous blood was collected. LncRNA HOTAIR/Galectin-3 expressions in serum were detected. Subjects were assigned into HOTAIR/Glactin-3 high/low expression groups and their correlation with age, gender, BMI, tumor size, pathological stage, TSH, TPO-Ab, and TG-Ab in PTC was analyzed. Receiver operating characteristic (ROC) curve was conducted on diagnostic efficacy of HOTAIR or/and Galectin-3. The difference of area under the curve (AUC) was compared and analyzed.HOTAIR and Glactin-3 were higher in PTC group and correlated with tumor pathological stage. Higher HOTAIR/Glactin-3 expression indicated a more advanced TNM stage. LncRNA HOTAIR was positively correlated with TPO-Ab and TG-Ab. AUC of HOTAIR for PTC diagnosis was 0.895, with 96.00% specificity and 80.63% sensitivity. AUC of Glactin-3 for PTC diagnosis was 0.817, with 66.67% specificity and 78.75% sensitivity. AUC of HOTAIR combining with Glactin-3 for PTC diagnosis was 0.969 with 96.00% specificity and 87.50% sensitivity. AUC of lncRNA HOTAIR was higher than that of Glactin-3, while AUC of the combination was higher than that of lncRNA HOTAIR or Glactin-3.LncRNA HOTAIR and Glactin-3 were highly expressed in PTC. The combination detection of lncRNA HOTAIR/Glactin-3 had higher diagnostic efficiency on the differential diagnosis of benign thyroid tumor and PTC.© 2021 Li et al.