The present study evaluated the clinical efficacy of chemotherapy in combination with cytokine-induced killer (CIK) biotherapy compared to the chemotherapy alone. Fifty-nine advanced non-small cell lung cancer (NSCLC) patients were randomly divided into two groups, group A (chemotherapy alone, including docetaxel 75 mg/m2, day 1; cisplatin, 25 mg/m2, days 1-4, tri-weekly) and group B (chemotherapy plus CIK cell transfusion). Autologous CIK cells were induced from the patients'peripheral mononuclear cells in vitro and separated by cytometry and then transfused back the patients. The host cellular immune function, clinical curative effects and quality of life (QOL) were examined and were compared between the two groups. The host immune function was enhanced and QOL was improved in the patients treated by chemotherapy plus CIK biotherapy compared to the patients treated by chemotherapy alone. The overall response rate (ORR) was 43.3% and 44.8% in groups A and B, respectively. The disease control rate (DCR) was higher in group B than in group A (89.7% vs. 65.5%, p = 0.030). The time to progression was 4.67 months (95% CI 3.98-6.02 months) in group A and 6.65 months (95% CI 4.70-7.30 months) in group B and the median survival time was 11.0 months (95% CI 7.88-14.1 months) in group A and 15.0 months (95% CI 11.04-18.96 months) in group B. Compared to patients in group A, the patients in group B had significantly longer progression-free survival (p = 0.042) and overall survival (p = 0.029). No severe side-effects occurred in the CIK cell transfusion patients. It was concluded that chemotherapy plus CIK cells has potential benefits compared to chemotherapy alone in patients suffering from advanced NSCLC and autologous CIK cell transfusion has no obvious side-effects.