Ubiquitin Specific Peptidase 53 Inhibits The Occurrence And Development Of Clear Cell Renal Cell Carcinoma Through NF-κB Pathway Inactivation

Background: Renal carcinoma is the most common type of kidney cancer,which arises from renal parenchymal urinary epithelial system and accounts for about 3% of adult malignancies.Clear cell renal cell carcinoma(cc RCC)is a type of RCC and occupies 80% of RCC.More than 140,000 people die from renal carcinoma each year,and nearly 30% of patients will still relapse and metastasize after undergoing local tumor resection.The root cause of renal carcinoma is difficult to diagnose and cure their pathogenesis and molecular mechanism is unclear,so it is particularly important to explore the molecular mechanism of renal carcinoma and find safe and effective biological targets.Studies have shown that E3 ubiquitin ligase VHL biallelic inactivation accounts for 50%-80% in kidney cancer patients,suggesting that the regulation of ubiquitination and deubiquitination is crucial in kidney cancer.Ubiquitin specific peptidase 53(USP53)belongs to the family of deubiquitinating enzymes.It has been reported that USP53 is associated with cholestatic liver disease,obesity,and Cantu syndrome in children,but its role in tumors still needs further study.Methods: We conducted differential genetic analysis of cc RCC in the GEO and TCGA databases through bioinformatics,and found that USP53 is related to the occurrence and development of cc RCC.We also constructed USP53 overexpression and knockdown 786-O and Caki-1 cells by means of lentivirus infection.And through CCK-8,EDU,Brd U cell proliferation experiments and scratches,Transwell cell transfer experiments to explore its potential biological function.Then we further explored the molecular mechanism of USP53 influencing the occurrence and development of cc RCC through RNA sequencing analysis and Western-blot experiments.Finally,we verified the effect of USP53 on cc RCC in the in vivo environment through nude mice subcutaneous tumorigenesis experiments.Results: Differential gene analysis of cc RCC showed that the gene expression of USP53 in cc RCC tissue was lower than that in normal tissue,and it was significantly negatively correlated with clinical prognosis.In the tumor grade of cc RCC,the expression of USP53 decreases as the malignant degree of cc RCC increases.Cell proliferation and metastasis experiments showed that overexpression of USP53 caused significant inhibition of cc RCC cell proliferation and metastasis;after USP53 knockdown,the proliferation and metastasis ability of cc RCC cells was significantly enhanced.RNA sequencing analysis showed that overexpression of USP53 caused the expression of a large number of inflammatory genes in the NF-κB signaling pathway to decrease.Western-blot results showed that after overexpression of USP53,the phosphorylation levels of IKKβ and P65 decreased,and the increased expression of IκBα resulted in the inhibition of NF-κB pathway activity;When USP53 was knocked down,the phosphorylation levels of IKKβ and P65 increased,the expression of IκBαdecreased,and the activity of this pathway increased.The experimental results of subcutaneous tumor formation in nude mice showed that the knockout of USP53 could significantly promote the growth of cc RCC.Conclusions: USP53 is probably regulated by the NF-κB pathway to regulate the occurrence and development of cc RCC.Our research reveals that USP53 may be a regulator of the NF-κB pathway and play a key role in the development of cc RCC.In addition,targeting USP53 may be a new strategy for the treatment of cc RCC.