The purpose of this study was to explore the possible involvement of SV40 polyomavirus in the development of renal cell carcinoma (RCC) in patients from the province of Al-Najaf. The study analyzed 75 paraffin-embedded block tissues of RCC, collected from archives of AL-Sader Medical City, and some private histopathology laboratories in Najaf governorate. The patients included 45 males and 30 females, aged between 22 and 70 years. The study used advanced scientific techniques, including Polymerase Chain Reaction (PCR) and immunohistochemistry (IHC), to detect the presence of SV40 and evaluate the expression state of Cyclin-Dependent Kinase Regulators (KAP or cyclin-dependent kinase inhibitor 3 (CDKN3) & Cyclin E1 markers). Hematoxylin and Eosin staining was used for diagnosing RCC. The study found that RCC is associated with the dysregulation of Cyclin-Dependent Kinase regulators (CDK), caused by the SV40 polyomavirus. The results of the IHC analysis showed an increased positive percentage for KAP or CDKN3 marker and a decreased positive percentage of Cyclin E1 marker. Additionally, the clear cell type was found to be the most common, accounting for 56% of the cases, while grade I was the most prevalent, representing 41.3% of the cases. Tumor stage type I was found to be higher, with 25 cases. PCR detected the presence of SV40 in 20 cases, accounting for 26.7% of the total cases studied. The study concluded that the Simian Virus 40 (SV40), particularly its Large T Antigen (Tag), affects CDK regulators and disrupts the delicate equilibrium of cell cycle regulation systems. Therefore, the study suggests a possible link between the development of renal cell carcinoma and the SV40 polyomavirus. The study recommends routine testing for the detection of RCC using PCR and IHC methods.