Breast cancer (BC) is the most common cancer among women in the world. The human multidrug resistance 1 gene (MDR1) is potentially an important gene influencing the susceptibility to breast cancer. This study aimed to assess the association of MDR1 genetic polymorphisms with the susceptibility to BC. Overall, 353 BC patients and 360 cancer-free controls were enrolled. The clinical characteristics were summarized by questionnaires. The c.1564A > T genetic polymorphism was genotyped using created restriction site-polymerase chain reaction method. We found that no significant differences in the genotypic and allelic frequencies between BC patients and cancer-free controls. Furthermore, the distribution of BC patients' risk factors was not significantly different among AA, AT, and TT genotypes. Our findings indicate that the c.1564A > T genetic polymorphism is not significantly associated with the susceptibility to BC in Chinese Han populations.