Abstract: Objective To investigate the mutation of echinoderm microtubule associated protein like 4-anaplastic lymphoma kinase（EML4-ALK） fusion gene in non-small cell lung cancer（NSCLC），and analyze its mutation with clinicopathologic characteristics and prognosis. Methods Amplification refractory mutation system was used to detect 9 mutations of EML4-ALK fusion gene and 4 mutations including exon 18，19，20 and 21 of epidermal growth factor receptor（EGFR）in 26 patients with NSCLC. The relationship between EML4-ALK mutations and clinicopathologic characters，overall survival（OS）was further analyzed. Results There were 6 patients with EML4-ALK fusion gene mutations out of 26 patients. EML4-ALK fusion gene mutations were not related to pathological types，gender，specimen types and serum carcinoembryonic antigen，but with smoking，age，clinical stage，EGFR mutations and histological differentiation. The median OS of patients with EML4-ALK fusion gene mutations was 9 months，while those without mutations were 12 months（P=0.460）. Conclusion EML4-ALK fusion gene mutations are often found in NSCLC patients with younger age，no history of smoking or a small amount of smoking，later clinical stage，poor histological differentiation and without EGFR mutation.