The regulatory relationship between silent information regulator 2 (SIRT2) and glucose 6-phosphate dehydrogenase (G6PD) in clear cell renal cell carcinoma (ccRCC) is still unclear. The present study aimed to explore the function of SIRT2 and its regulatory effect on G6PD in ccRCC. The Cancer Genome Atlas data mining of SIRT2 was first analyzed. RT-qPCR and western blotting analyses were used to assess the mRNAs and proteins expressions levels respectively. The cell viability assay, colony formation assay, EdU staining, cell cycle assay, cell apoptosis assay, and TUNEL assay were used to investigate the roles of SIRT2 in ccRCC proliferation and apoptosis. The co-immunoprecipitation (Co-IP) assay was performed to analyze the association of SIRT2 with G6PD in ccRCC cells. Quantitative Co-IP assay was used to detect the levels of G6PD ubiquitination and small ubiquitin-related modifier 1 (SUMO1). A in vivo experiment was also performed to confirm in vitro findings. The results indicated that SIRT2 promoted ccRCC proliferation and inhibited apoptosis by regulating cell cycle and apoptosis related proteins. SIRT2 interacted with G6PD, facilitated its activity through deacetylation and increased its stability by reducing its ubiquitination and enhancing its SUMO1 modification. SIRT2 also promoted ccRCC tumor development in vivo. Taken together, the present study demonstrated that SIRT2 promoted ccRCC progression by increasing G6PD activity and stability, and it could be a potential new diagnostic and therapeutic target for ccRCC.This article is protected by copyright. All rights reserved.