This study aimed to determine the impact of vitamin D deficiency on Pap smear test results in women in the postmenopausal period. This cross-sectional study included 394 women who applied to İstanbul Medeniyet University, Göztepe Training and Research Hospital gynecology and obstetric clinic for routine Pap smear tests. This study was conducted between May 2019 and December 2020. Cases were divided into severe deficiency (<12 ng/ml), moderate deficiency (12-20 ng/ml), deficiency (21-30 ng/ml), and sufficiency (>30 ng/ml) according to the vitamin D levels measured in the postmenopausal period within the scope of check-up examinations. Pap smear test results compared with vitamin D levels. The mean age of the study group was 48.70±3.46 years. There was a statistically significant association between vitamin D values and Pap smear test results (p<0.05). Vitamin D greater than 20 ng/ml had a significant relationship with the normal cytology results. There was a statistically significant association between vitamin D lower than 20 ng/ml and cervicitis cytology results. The results showed that the serum vitamin D and the Pap smear test results had a significant association. It is recommended that physicians prescribe the use of vitamin D supplements in women in the high-risk group of cervical cancer.