Background: Thyroid cancer (TC) is the most common endocrine malignancy in the world. Previous studies that link the levels of thyroid-stimulating hormone (TSH) and 25-hydroxyvitamin D (25(OH) D) to TC have been relatively inconsistent. Our goal is to assess alterations in serum levels of TSH and 25(OH)D between TC patients and non-toxic goiter patients and to examine the association between TSH and 25(OH) D levels in Saudi patients. Methods: A retrospective analysis of serum hormone fluctuations in primary TC (n= 162) and non-toxic goiter (n=98) cases. Results: TSH was significantly higher in TC patients than in non-toxic goiter patients (P<0.05). When gender was taken into consideration, the difference in females remained significant but was eliminated in males. Conversely, there was an insignificant difference in 25(OH) D between the two groups (P>0.05). Spearman rank correlation analysis demonstrated a statistically insignificant association between TSH and 25(OH) D (r=0.011), TSH and (r=0.005), or 25 (OH)D and body mass index BMI (r=0.159) in TC patients. Conclusion: TSH level is associated with TC in female Saudi patients, independent of 25(OH) D or obesity.