Various studies have demonstrated that the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene polymorphism contributes to the risk of prostate cancer, while other studies have provided conflicting findings. In the present study, we carried out a comprehensive meta-analysis with the aim of determining whether there is a significant association of the MTHFR gene A1298C polymorphism with the susceptibility of prostate cancer. Studies on the MTHFR gene A1298C polymorphism and prostate cancer were retrieved using the electronic PubMed database without any restriction on language through Aug 21, 2011. Data were abstracted by a standardized protocol. Crude odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to estimate the strength of association. The analyses were conducted with Review Manager software version 4.2. Nine case-control studies were identified, including 2,723 prostate cancer patients and 3,442 controls. Overall, no significant associations were found between the MTHFR gene A1298C polymorphism and prostate cancer (codominant models: CC vs. AA, OR=1.03, 95% CI 0.79-1.34, P=0.84; AC vs. AA, OR=1.04, 95% CI 0.93-1.16, P=0.46; dominant model: AC + CC vs. AA, OR=1.04, 95% CI 0.94-1.15, P=0.48; recessive model: CC vs. AC + AA, OR=1.02, 95% CI 0.76-1.35, P=0.91; allele model: C vs. A, OR=1.04, 95% CI 0.90-1.19, P=0.61). Similarly, in the subgroup analyses by DNA source, ethnicity, control source, pathological stage and Hardy-Weinberg equilibrium, no significant associations were observed. Our meta-analysis suggests that the MTHFR gene A1298C polymorphism is not associated with the risk of prostate cancer.