Abstract: Objective To analyze the role of ultrasound-guided biopsy in diagnosis of non-small cell lung cancer (NSCLC)patients with cervical lymph node enlargement. Methods Clinical data about 64 NSCLC patients with cervical or supraclavicular lymph node enlargement admitted to Chinese PLA General Hospital from May 2011 to December 2013 were retrospectively analyzed. Biopsy findings and malignancy of primary foci were compared. The relation between lymph node size, blood flow, boundaries between cortex and medulla, lymph door structure, benign or malignant lesion was analyzed. Results The incidence of malignant cervical lymph nodes was 81.2%. Of the 64 patients, 30 were diagnosed with adenocarcinoma, 18 were diagnosed with squamous cell carcinoma, 4 were diagnosed with lymphoma, and 12 were diagnosed with infammatory hyperplasia. The average diameter of cancerous lymph nodes was 2.1±0.6 cm. The positive blood fow signals accounted for 57%, unclear boundaries between cortex and medulla accounted for 43%, and lymph door-free structure accounted for 61% (P< 0.01). Conclusion Cervical and supraclavicular ultrasonography plays an important role in diagnosis of preoperative lymph node metastasis and assessment of the postoperative condition in NSCLC patients.