ObjectiveTo compare the surgical efficacy of Da-Vinci robot-assisted thoracic surgery (RATS) and video-assisted thoracic surgery (VATS) for non-small cell lung cancer (NSCLC). MethodsOnline databases including PubMed, EMbase, Web of Science, The Cochrane Library, CNKI, Wanfang, VIP and CBM from inception to 18 February, 2022 were searched by two researchers independently. The references of related studies were also searched to re-enroll the potential studies. The quality of the studies was evaluated with Newcastle-Ottawa Scale (NOS). The meta-analysis was performed by RevMan 5.3. Results A total of 43 studies including 33 089 patients were enrolled in the final study. The NOS scores of the included studies were ≥6 points. The results of meta-analysis showed that the operation time was longer [MD=8.50, 95%CI (1.59, 15.41), P=0.020], the blood loss was less [MD=−46.58, 95%CI (−62.86, −30.29), P＜0.001], the dissected lymph nodes stations were more [MD=0.67, 95%CI (0.40, 0.93), P＜0.001], the dissected lymph nodes were more [MD=2.39, 95%CI (1.43, 3.36), P＜0.001], the conversion rate was lower [OR=0.52, 95%CI (0.46, 0.59), P＜0.001], the time of chest tube drainage was shorter [MD=−0.35, 95%CI (−0.58, −0.11), P=0.004], the length of hospital stay was shorter [MD=−0.32, 95%CI (−0.45, −0.19), P＜0.001], and the recurrence rate was lower [OR=0.51, 95%CI (0.36, 0.72), P＜0.001] in the RATS group than those in the VATS group. The rate of overall postoperative complications [OR=0.95, 95%CI (0.89, 1.01), P=0.110] and postoperative mortality rate [OR=0.85, 95%CI (0.62, 1.16), P=0.300] were not significantly different between the two groups. ConclusionCompared with VATS, although RATS prolongs the operation time, it does not increase the incidence of postoperative complications and mortality rates. Moreover, RATS can dissect more lymph nodes, effectively control intraoperative bleeding, shorten the duration of chest drainage tube indwelling and shorten the postoperative hospital stay to a certain extent.