Non-small cell lung cancer (NSCLC) represents the most common cause of cancer mortality worldwide and counts for the greatest number of deaths from lung cancer in both men and women over age 60. Pemetrexed is a multi-targeted anti-metabolite and has shown comparable activity for Caucasian patients with advanced NSCLC. In this single-center retrospective study, we analyzed the outcome in 75 Chinese non-squamous NSCLC patients treated with pemetrexed as first-line therapy and assessed its efficacy and tolerability. The overall response rate was 9.3% (7/75) with 7 partial response s and no complete response. There were 44 (58.7%) stable diseases and 24 progressive diseases (32.0%). The median progression-free survival (PFS) was 6.79months (95% CI 5.69–7.90months), and the median overall survival (OS) was 11.67months (95% CI 9.98–13.36months). Good performance status and negative smoke history predicted better PFS and OS. Most side effects were generally mild and well tolerated. Taken together, pemetrexed was safe and effective in Chinese patients with non-squamous NSCLC. Pemetrexed alone or in combination with other efficacious drugs may serve as first-line therapy for this disease.