Abstract: Objective To explore the expression of CD166 and transcription factor MALAT1 gene in the occurrence and development of non-small cell lung cancer (NSCLC). Methods 100 cases of patients undergoing lung cancer radical resection with pathological diagnosed NSCLC were chosen as the research object. The selected fresh NSCLC tissues were used as observation group, tissue adjacent to carcinoma as the control group. The CD166 and MALAT1 mRNA expression in both groups were determined by immunohistochemical and RT-PCR. Results The observation group’s CD166 positive expression rate was significantly higher than the control group (P < 0.05). Observation group MALAT1 expression was significantly higher than the control group (P<0.05). The MALAT1 and CD166 expression were related with pathological grade (P < 0.05). Conclusion The CD166 and MALAT1 gene transcription factor might have important role in the process of occurrence and development of NSCLC and might have certain value in the diagnosis of NSCLC.