OBJECTIVE Nonsyndromic cleft lip with or without palate (NSCL/P) represents a complex condition caused by genetic and environmental factors. The aim of this study was to investigate the relationship between the EPHA3 polymorphisms and NSCL/P. METHODS To investigate the relationship between five EPHA3 single nucleotide polymorphisms (SNPs) and NSCL/P, we selected 180 affected patients and 167 normal controls from the Chinese Han Population. EPHA3 SNPs (rs7650466, rs1398197, rs17801309, rs1054750, and rs7632427) were genotyped using the SNaPshot technique; bioinformatic analyses were performed to determine if any of them were potentially functional SNPs. RESULTS The rs7650466 T allele was associated with the incidence of NSCL/P (OR, 0.211; 95% CI, 0.131-0.338; adjusted p = 4.881 × 10-10) and cleft lip with or without palate (CL/P) (OR, 0.176; 95% CI, 0.104-0.297; adjusted p = 3.617 × 10-10), as well as with protective and dominant effects in both conditions. The rs7650466 T allele could be associated with reduced risk of the malformation. In a bioinformatics analysis, we found potential matching sites (miR-1255a, miR-125a-3p, miR-143, and miR-552) for rs7650466 and preliminarily analyzed its potential function. CONCLUSIONS Collectively, our data suggest that the EPHA3 rs7650466 polymorphism confers genetic risk for NSCL/P in the Chinese Han Population. Furthermore, rs7650466 is associated with CL/P incidence in stratified analyses, but not with cleft palate only.