ARRDC3 regulates the targeted therapy sensitivity of clear cell renal cell carcinoma by promoting AXL degradation

ABSTRACT

AXL plays crucial roles in the tumorigenesis, progression, and drug resistance of neoplasms; however, the mechanisms associated with AXL overexpression in tumors remain largely unknown. In this study, to investigate these molecular mechanisms, wildtype and mutant proteins of arrestin domain-containing protein 3 (ARRDC3) and AXL were expressed, and co-immunoprecipitation analyses were performed. ARRDC3-deficient cells generated using the CRISPR-Cas9 system were treated with different concentrations of the tyrosine kinase inhibitor sunitinib and subjected to cell biological, molecular, and pharmacological experiments. Furthermore, immunohistochemistry was used to analyze the correlation between ARRDC3 and AXL protein expressions in renal cancer tissue specimens. The experimental results demonstrated that ARRDC3 interacts with AXL to promote AXL ubiquitination and degradation, followed by the negative regulation of downstream signaling mechanisms, including the phosphorylation of protein kinase B and extracellular signal-regulated kinase. Notably, ARRDC3 deficiency decreased the sunitinib sensitivity of clear cell renal cell carcinoma (ccRCC) cells in a manner dependent on the regulation of AXL stability. Overall, our results suggest that ARRDC3 is a negative regulator of AXL and can serve as a novel predictor of sunitinib therapeutic response in patients with ccRCC.

KEYWORDS:

• ARRDC3
• AXL
• clear cell renal cell carcinoma
• sunitinib
• ubiquitination

Abbreviations

ARRDC3	=	arrestin domain-containing protein 3;
ccRCC	=	clear cell renal cell carcinoma;
GAPDH	=	glyceraldehyde 3-phosphate dehydrogenase;
Ig	=	immunoglobulin;
IP	=	Immunoprecipitation;
ITGβ4	=	integrin β4;
PAR1	=	protease-activated receptor 1;
RCC	=	Renal cell carcinoma;
WB	=	western blotting;
YAP1	=	yes-associated protein-1.

Acknowledgements

This research was encouraged and supported by the National Natural Science Foundation of China, grants 81372753 (J.F.) and 82172612 (J.F.).

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Author contributions

All authors have contributed significantly to the manuscript and agree with its content. Mulin Chen performed most of the experiments and the statistical analysis; Bingde Yin provided technical support for the assays; Yao Liu, Mingzi Li, and Suqin Shen offered advice for improving the experiments; Jiaxue Wu designed the experiments and wrote the manuscript; Weiguo Li provided guidance on study direction, and Jie Fan conceived the study and contributed to the completion of the manuscript.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/15384101.2024.2308411

Correction Statement

This article has been corrected with minor changes. These changes do not impact the academic content of the article.

Additional information

Funding

The work was supported by the National Natural Science Foundation of China [82172612]; National Natural Science Foundation of China [81372753].