Li Hongsheng, Ning Yao, Yu Junjie, Chen Yiju, He Qiang, Jin Juan
Department of Laboratory Medicine, The Second Affiliated Hospital of Jiaxing University, Jiaxing, China.
Department of Nephrology, Zhejiang Key Laboratory of Research and Translation for Kidney Deficiency-Stasis-Turbidity Disease, Zhejiang-Macau International Joint Laboratory of Integrated Traditional Chinese and Western Medicine for Nephrology and Immunology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, Zhejiang, China.
Cancer Biol Ther. 2025 Dec;26(1):2558402. doi: 10.1080/15384047.2025.2558402. Epub 2025 Sep 15.
Ubiquitin-specific protease 7 (USP7) involves in various human cancers due to its capacity for binding and stabilizing specific target proteins through deubiquitylation, but its roles in clear cell renal cell carcinoma (ccRCC) development remains unknown. This study aimed to determine the role of USP7 in the pyroptosis mechanism in ccRCC, thereby providing novel anti-ccRCC strategies. Bioinformatics analysis was conducted to explore the expression of USP7 and TRIP12 in ccRCC patients and their association with patient overall survival. qRT-PCR, western blotting, and ELISA were used to determine the levels of USP7, TRIP12, pyroptosis-related factors. Cell viability, invasion, pyroptosis, and proliferation were evaluated using CCK-8, Transwell, flow cytometry, and immunohistochemistry assays. The direct interaction between USP7 and TRIP12 was validated by co-immunoprecipitation (CO-IP). We found downregulated USP7 in ccRCC tissues, which was related to the shorter patient overall survival (OS). Significantly, USP7 was also decreased in ccRCC cells. oe-USP7 (USP7 overexpression) inhibited ccRCC cell viability, migration, invasion, and enhanced pyroptosis. The caspase-1 specific inhibitor, VX-765, partially abolished the anti-viability, and pro-pyroptosis effects of oe-USP7, indicating USP7 overexpression prevented the malignant phenotype of ccRCC cells by enhancing caspase-1 dependent pyroptosis. Similarly, the shorter patient OS was indicated to be associated with reduced TRIP12 in ccRCC tissues. Besides, oe-USP7 increased TRIP12 expression in ccRCC cells by deubiquitinating TRIP12, while sh-TRIP12 eliminated the biological functions of oe-USP7. The similar effects of oe-USP7 on ccRCC development were found in ccRCC mice. USP7 mediated TRIP12 deubiquitination inhibited ccRCC progression by enhancing pyroptosis.
泛素特异性蛋白酶7（USP7）因其具有通过去泛素化结合并稳定特定靶蛋白的能力而参与多种人类癌症，但它在透明细胞肾细胞癌（ccRCC）发展中的作用尚不清楚。本研究旨在确定USP7在ccRCC细胞焦亡机制中的作用，从而提供新的抗ccRCC策略。进行生物信息学分析以探索USP7和TRIP12在ccRCC患者中的表达及其与患者总生存期的关联。采用qRT-PCR、蛋白质免疫印迹法和酶联免疫吸附测定法来测定USP7、TRIP12、焦亡相关因子的水平。使用CCK-8法、Transwell小室法、流式细胞术和免疫组织化学分析法评估细胞活力、侵袭能力、焦亡和增殖情况。通过免疫共沉淀（CO-IP）验证USP7与TRIP12之间的直接相互作用。我们发现ccRCC组织中USP7表达下调，这与患者较短的总生存期（OS）相关。值得注意的是，ccRCC细胞中USP7也减少。过表达USP7（oe-USP7）抑制ccRCC细胞活力、迁移、侵袭，并增强焦亡。半胱天冬酶-1特异性抑制剂VX-765部分消除了oe-USP7的抗生存能力和促焦亡作用，表明USP7过表达通过增强半胱天冬酶-1依赖性焦亡来阻止ccRCC细胞的恶性表型。同样，ccRCC组织中患者较短的OS表明与TRIP12减少有关。此外，oe-USP7通过使TRIP12去泛素化增加ccRCC细胞中TRIP12的表达，而短发夹RNA干扰TRIP12（sh-TRIP12）消除了oe-USP7的生物学功能。在ccRCC小鼠中发现oe-USP7对ccRCC发展有类似作用。USP7介导的TRIP12去泛素化通过增强焦亡抑制ccRCC进展。
