Despite the promise of targeted tyrosine kinase inhibitors (TKIs), such as sunitinib, in the extension of survival time in patients with clear cell renal cell carcinoma (ccRCC) progression or metastasis, the patients eventually succumb to inevitable drug resistance. Protein degradation executed by the ubiquitin-dependent proteasome system played an important role in determining the sensitivity of ccRCC to sunitinib. Here, we applied the bioinformatic analysis to identify that E3 ligase RBCK1 was elevated in the sunitinib-resistant renal cancer cell lines or patient specimens. The subsequent in vitro or in vivo studies demonstrated that RBCK1 contributed to decreasing the sensitivity of ccRCC to sunitinib. Then, we showed that inhibition of RBCK1 inactivated the AKT and MAPK signaling pathways, which might be one of the main reasons why RBCK1 induces sunitinib resistance in ccRCC cells. Mechanistically, our results indicated that RBCK1 promotes the degradation of ANKRD35 and that ANKRD35 destabilizes MITD1 by binding with SUMO2 in ccRCC cells. In addition, we showed that the RBCK1-ANKRD35-MITD1-ANXA1 axis regulates the phosphorylation of AKT and ERK and contributes to the dysregulation of sunitinib in ccRCC cells. Therefore, we identified a novel mechanism for regulating the sensitivity of sunitinib in ccRCC. Therefore, we elucidated a novel mechanism by which RBCK1 regulates sunitinib sensitivity in ccRCC.
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The datasets used and/or analyzed during the current study are available from the corresponding authors on reasonable request. The original western blot data has been submitted to the figshare repository (https://doi.org/10.6084/m9.figshare.21557733). The sequencing data was deposited in the GEO public dataset (GSE214635).
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This work was supported by grants from the Chinese National Natural Science Foundation Grant No. 82073321 (XJ), 82272910 (XJ) and 82103341 (WX). Excellent Youth Foundation of Hunan Scientific Committee (Grant No. 2022JJ10092, XJ). Hunan leading program for science and technology innovation of high technology industries (Grant No. 2022GK4020, XJ). Central South University Innovation-Driven Research Programme (Grant No. 2023CXQD059, XJ). Natural Science Foundation of Hunan Province of China (Grant No. 2022JJ30870 (LZ), 2022JJ40719 (WX)).
The authors declare no competing interests.
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The study was conducted in accordance with the principles of the Declaration of Helsinki principles (Approved no. 2021068). It was approved by the Animal Use and Care Committees at the Second Xiangya hospital, Central South University.
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Wang, Y., Peng, M., Zhong, Y. et al. The E3 ligase RBCK1 reduces the sensitivity of ccRCC to sunitinib through the ANKRD35-MITD1-ANXA1 axis. Oncogene 42, 952–966 (2023). https://doi.org/10.1038/s41388-023-02613-w