Objective To systematically evaluate the effectiveness and safety of disodium cantharidinate and vitamin B6 injection plus chemotherapy compared with chemotherapy alone, in the treatment of non-small cell lung cancer (NSCLC). Methods The Cochrane Library (Issue 1, 2011), MEDLINE (1966 to November 2011), EMbase (1984 to November 2011), CBM (1978 to November 2011), CNKI (1995 to November 2011) and VIP (1989 to November 2011) were searched electronically, and the randomized controlled trials (RCTs) about disodium cantharidinate and vitamin B6 injection plus chemotherapy for NSCLC were included. The quality of the included studies was assessed and crosschecked by two reviewers independently, and meta-analyses were performed for homogeneous studies by using Cochrane Collaboration’s RevMan 5.1 software. Results Eight RCTs involving 539 patients met inclusion criteria were included in meta-analyses. The quality of all studies was in Grade B. The results of meta-analyses showed that disodium cantharidinate and vitamin B6 injection plus chemotherapy, compared with chemotherapy alone, could increase effective rate (RR=1.32, 95%CI 1.07 to 1.62) and clinical benefit rate (RR=1.24, 95%CI 1.12 to 1.37), improve quality of life (RR=2.23, 95%CI 1.55 to 3.19) and clinical symptoms (RR=1.55, 95%CI 1.24 to 1.95), increase body weight (RR=2.72, 95%CI 1.74 to 4.25), and decrease bone marrow suppression (leucocyte reduction rate) (RR=0.36, 95%CI 0.21 to 0.61). Conclusion The evidence available indicates that the treatment regimen of disodium cantharidinate and vitamin B6 injection plus chemotherapy is superior to chemotherapy alone in increasing effects and decreasing toxicity for the patients with NSCLC. More high-quality and multi-center RCTs with larger sample and longer follow-up are proposed.