Abstract
Objective: Renal cell carcinoma (RCC) is a prevalent malignant tumor of the kidney that has considerable heterogeneity and intricate molecular pathways. Incidence and death rates associated with kidney cancer have been increasing steadily. Consequently, investigating its molecular basis and identifying new biomarkers for targeted therapies have become important. Ubiquitin C-terminal hydrolase L5 (UCHL5) has an effect on tumor initiation and progression. This study aims to explore the role of UCHL5 in promoting RCC development and identify related molecular mechanisms. Material and methods: Quantitative real-time polymerase chain reaction and Western blot (WB) analyses were employed to measure UCHL5 expression. Stable UCHL5 overexpression and knockdown cell lines were generated using lentiviral transfection in 786-O cells. Colony formation and 5-ethynyl-2'-deoxyuridine assays were used to assess cell proliferation. Cell migration was evaluated using scratch assays, while invasion capacity was determined by Transwell assays. Commercial kits were utilized to quantify glucose consumption and lactate production. WB was applied to detect proteins related to glycolysis and components of the Phosphoinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. The function of UCHL5 in RCC cells was validated in vivo through an animal tumor model. Results: The expression of UCHL5 increased in RCC cell lines (P < 0.01). Silencing UCHL5 significantly impaired the malignant behavior of RCC cells (P < 0.01), whereas its overexpression promoted this cellular behavior (P < 0.05). Reducing UCHL5 levels decreased glucose uptake, lactate secretion, and protein expression involved in glycolysis, while overexpression enhanced glycolytic activity in RCC cells (P < 0.01). Knockdown of UCHL5 diminished the phosphorylation of PI3K/AKT/mTOR pathway proteins (P < 0.05), whereas the opposite effect was observed upon overexpression (P < 0.01). Treatment with 740Y-P effectively reversed the suppression of glycolysis, proliferation, and metastasis caused by UCHL5 knockdown (P < 0.01). Conversely, LY294002 inhibited the glycolytic enhancement and aggressive phenotypes induced by UCHL5 overexpression (P < 0.01). Animal experiments further confirmed that UCHL5 downregulation suppressed RCC growth through the PI3K/AKT/ mTOR pathway (P < 0.01). Conclusion: UCHL5 activates the PI3K/AKT/mTOR cascade, which enhances the glycolysis of RCC cells and promotes the development of renal cancer. This study provides insights into the molecular mechanisms underlying the oncogenic role of UCHL5 in RCC.