The CXCL12-3' G801A transition (rs1801157) has been associated with the incidence of breast cancer. However, the contribution of CXCL12-3' G801A polymorphisms in breast cancer development and progression has been controversial. Therefore, we examined the incidence of CXCL12-3' G801A polymorphic variants in patients with breast cancer (n = 193) and controls (n = 199) in a Polish cohort. We observed a trend of slightly increased presence of CXCL12-3' AA and GA genotypes and CXCL12-3'A allele frequency in patients with breast cancer compared with healthy individuals. However, these differences between cases and controls were not statistically significant. Odds ratio (OR) for patients with breast cancer and the CXCL12-3' A/A genotype was 1.898 (95% confidence interval [CI] = 0.6242-5.770, p = 0.2866) and OR of the CXCL12-3' A/A and A/G genotypes was 1.229 (95% CI = 0.8082-1.868, p = 0.3395). OR for the CXCL12-3'A allele frequency was 1.249 (95% CI = 0.8716-1.789, p = 0.2352). Our investigation did not support the CXCL12-3'A gene variant as a risk factor for breast cancer incidence in a sample of the Polish population.