Simple SummaryThe current outcomes for early diagnosis and late treatment of clear cell renal cell carcinoma (ccRCC) are unsatisfactory. This study aimed to determine the diagnostic and prognostic value of Netrin family genes for ccRCC and to explore new therapeutic targets. We systematically analyzed the expression profile of Netrin family genes in ccRCC using a bioinformatics approach and identified three genes, including NTN4, as potential biomarkers for ccRCC prognosis. In vitro and in vivo experiments confirmed that NTN4 regulates the growth and invasion of ccRCC cells by inhibiting the Wnt/β-catenin signaling pathway. Targeting this pathway through NTN4 may be a promising strategy for ccRCC treatment. The results of this study may have important implications for developing personalized cancer treatment strategies and identifying new biomarkers for ccRCC.AbstractClear cell renal cell carcinoma (ccRCC, or KIRC) is the most common type of kidney cancer, originating within the renal cortex. The current outcomes for early diagnosis and late treatment of ccRCC are unsatisfactory. Therefore, it is important to explore tumor biomarkers and therapeutic opportunities for ccRCC. In this study, we used bioinformatics methods to systematically evaluate the expression and prognostic value of Netrin family genes in ccRCC. Through our analysis, three potential biomarkers for ccRCC were identified, namely NTNG1, NTNG2, and NTN4. Moreover, we performed in vitro and in vivo experiments to explore the possible biological roles of NTN4 and found that NTN4 could regulate ccRCC development through Wnt/β-catenin signaling. We elucidate the molecular mechanism by which NTN4 modulates β-catenin expression and nuclear translocation to inhibit ccRCC progression, providing a new theoretical basis for developing therapeutic targets for ccRCC. Thus, we suggest that Netrin-related studies may offer new directions for the diagnosis, treatment, and prognosis of ccRCC patients.