Vitamin D has anticarcinogenic properties and acts through vitamin D receptor (VDR) to carry out its functions. This study explored the independent and combined effects of vitamin D statusand VDR genetic polymorphisms on breast cancer (BC) risk in Pakistani population.This case–control study recruited 100 cases with histologically confirmed breast cancer and 100 age-matched controls. Serum 25(OH) vitamin D level was measured using Enzyme-linked immunosorbent assay (ELISA) kit. VDR genotype was conducted for Apa I (rs7975232), Taq I (rs731236) and Fok I (rs2228570), using the tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR) and confirmation by Sanger sequencing. Logistic regression was usedto calculate odds ratio and 95% confidence interval after adjusting for various confounders.On average cases possessed a lower mean+Standard Deviation, serum vitamin D compared to control (cases: 15.1±9.5 ng/dl, vs controls: 19.6±9.3 ng/dl, pA polymorphism was significantly higher in BC patients than in control subjects (37% versus 25% respectively, adjusted OR= 2.1, 95% Cl: 1.09-4.1, p<0.05).A significant association exists between vitamin D deficiency and GA genotype of Taq I (rs731236) may be a susceptibility risk factor for BC development in the Pakistani population.