Uterine leiomyomas (ULs), commonly known as fibroids, are the most prevalent benign tumors affecting the smooth muscle of the uterus, impacting up to 60% of women of reproductive age. Emerging studies indicate an association between VDR polymorphisms and various cancers of the female reproductive system, including breast, ovarian, cervical, endometrial, uterine, and vaginal cancers. In relation to uterine fibroids, several gene polymorphisms exist, including those within the VDR gene. This study seeks to examine the influence of Fokl variants of the VDR gene—specifically focusing on susceptibility to ULs and their effects on VDR mRNA and serum vitamin D concentrations. A total of 200 participants, including 100 cases of UL and 100 controls matched for age and gender, underwent genotyping using TETRA ARMS PCR, followed by Sanger sequencing validation. Levels of VDR mRNA and vitamin D were also assessed through quantitative real-time PCR and ELISA methods, respectively. The association between this variant and leiomyomas was analyzed, along with clinico-pathological (obesity) associations. FokI exhibited a significant association with UL, especially with the CC genotype (odds ratio [OR]: 2.2; confidence interval [CI]: 1.00–4.93). VDR mRNA expression was found to be two times lower in UL patients (p < .001), along with decreased serum vitamin D levels (p < .0001). Correspondingly, homozygous genotypes of FokI were associated with lower serum vitamin D levels (p < .001). This research highlights the complex connection between VDR genetic variations, altered VDR functionality, and vitamin D metabolism in UL. Additional studies involving various populations are essential to confirm and generalize these results, potentially leading to tailored therapeutic approaches for vitamin D-associated disorders.