Objective:To investigate the expression and significance of cancer stem cell markers CD133 and TROP-2 in non-small cell lung carcinoma(NSCLC). Methods:The expression of CD133 and TROP-2 in NSCLC was detected by immunohistochemistry and double-labelling immunofluorescence. The location and the colocalization of the two proteins were observed under light microscope and confocal laser scanning microscope. Results:①The expression rate of CD133 and TROP-2 were 30.0% and 41.25% in 80 NSCLC tissues respectively,which were significantly higher than that in the tumor-adjacent normal tissues or benign lesion tissues(P < 0.01);②CD133-positive expression was significantly higher in poor differentiated tumor than that in the well differentiated tumor(P = 0.024);③ Positive expression of CD133 and TROP-2 in the lymph node metastasis group was significantly higher than that in the non-lymph node metastasis group(P < 0.05). The positive expression rate of TROP-2 increased with the increasing Tumor-Nodes-Metastasis (TNM) stage(P < 0.05). ④CD133 expression was correlated with TROP-2 expression (P < 0.001). And there was collocation between the CD133 and TROP-2 proteins. Conclusion:The expression CD133 and TROP-2 was associated with the invasion and metastasis of NSCLC. Their colocalization in NSCLC was detected preliminarily. CD133 and TROP-2 may be used as two important markers in judging the cancer stem cell of NSCLC.