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Objective: To investigate the expression and clinical significance of co-signal molecule B7-H3 in non-small cell lung cancer (NSCLC) cell lines and tissues. Methods: B7-H3 expression level was evaluated by RT-PCR and western blot in five NSCLC cell lines. Specimens from 121 participants were stained immunohistochemically. Staining was quantified using Image-Proplus software, and integrated optical density was measured on each section. Results: The constitutive expression of B7-H3 in all five cell lines was found. The expression of B7-H3 were higher than that in benign lesions and non-cancer tissues respectively (P<0.001). B7-H3 expression in well differentiated NSCLC was Inhigher than that in not well differentiated one (P<0.05).There was a significant association between expression of B7-H3 and lymph node metastasis. Conclusion: B7-H3 is overexpressed in NSCLC tissues and may be an important target for diagnosis or the therapy of the disease. [ABSTRACT FROM AUTHOR]