The association between MHTFR Glu429Val polymorphism and breast cancer risk are inconclusive. To derive a more precise estimation of the relationship, a meta-analysis was performed. A total of 29 studies including 8,649 cases and 18,672 controls were involved in this meta-analysis. Overall, no significant associations were found between MTHFR Glu429Ala polymorphism and breast cancer risk when all studies were pooled into the meta-analysis (Glu/Glu vs Ala/Ala: OR = 0.891, 95 % CI = 0.782-1.015; Glu/Ala vs Ala/Ala: OR = 0.874, 95 % CI = 0.760-1.006; dominant model: OR = 0.885, 95 % CI = 0.775-1.012; and recessive model: OR = 0.989, 95 % CI = 0.931-1.051). In the subgroup analysis by ethnicity, significantly elevated breast cancer risk was associated with Glu/Glu variant genotype in homozygote comparison and recessive genetic model (Glu/Glu vs Ala/Ala: OR = 0.78, 95 % CI = 0.63-0.97; Glu/Glu vs Glu/Ala: OR = 0.92, 95 % CI = 0.85-0.99; recessive model: OR = 0.91, 95 % CI = 0.85-0.97), while no significant associations were found for all comparison models in other ethnicity populations. In conclusion, this meta-analysis suggests that the MTHFR Glu429Ala polymorphism may be not associated with breast cancer development.