The aim of this study was to evaluate the efficacy of methotrexate (MTX) in the treatment of ankylosing spondylitis (AS). The literature on controlled clinical trials was searched from MEDLINE, EMBASE, OVID, and Cochrane Library databases up to November 2012. The quality of the studies included was evaluated publicly by two reviewers. A meta-analysis was conducted to the homogeneous studies using Cochrane systematic review. Three trials involving 116 patients compared treatment with MTX against placebo. No statistically significant differences (p < 0.05) were found in the primary outcome measures of withdrawal rate, bath ankylosing spondilitis active index (BASDAI), C-reactive protein (CRP), patient global assessment, and side effects such as nausea and vomiting. Two trials involving 142 patients compared treatment with MTX plus infliximab (IFX) against IFX alone in the effect of treatment of AS. No statistically significant differences (p < 0.05) were found in the primary outcome measures of ASAS20 and withdrawal rate. Thus, we should choose the right drugs based on the specific situation in clinical applications. Randomized controlled trials designed rationally and implemented strictly with multi-center, large sample size and enough follow-up time are needed in future research.