Lian Weibin, Hong Chengye, Chen Debo, Wang Chuan
Department of Breast Surgery, Fujian Medical University Union Hospital, Fuzhou, People's Republic of China.
Department of Breast Surgery, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, People's Republic of China.
Am J Physiol Cell Physiol. 2025 Feb 1;328(2):C355-C366. doi: 10.1152/ajpcell.00733.2024. Epub 2024 Dec 23.
Ubiquitin-specific protease 35 () was found to be involved in various tumor progression, but its role in breast cancer remains largely unknown. USP35 mRNA and protein expression in breast cancer tissues and cells were evaluated by quantitative real-time PCR and Western blot, respectively. Subsequently, flow cytometry and 5-ethynyl-2'-deoxyuridine labeling were used to evaluate breast cancer cell apoptosis and proliferation. Cellular glycolytic function was analyzed using the Seahorse assay and various kits. Furthermore, co-immunoprecipitation (Co-IP) and immunoprecipitation assays were utilized to validate the deubiquitylation mechanism of USP35. Finally, a subcutaneous human xenograft tumor model was established in nude mice to verify the effect of USP35 in vivo. By examining the clinical samples and cell lines, we found that USP35 expression was significantly upregulated in breast cancer. Further functional studies showed that knockdown USP35 expression inhibited cell proliferation and promoted apoptosis. In addition, knockdown of USP35 decreased phosphofructokinase1 (PFK-1) expression and was associated with lower extracellular acidification rate and oxygen consumption rate compared with sh-Control. Co-IP assays identified PFK-1 as a direct deubiquitiation target of USP35. Importantly, we demonstrated that PFK-1 is an essential mediator for USP35-induced cell proliferation and glycolysis in vitro and in vivo. This study identified that USP35 regulates the proliferation and glycolysis of breast cancer cells by mediating the ubiquitination level of PFK-1. The USP35/PFK-1 axis offers novel insight for the treatment of breast cancer. This study identified that USP35 regulates the proliferation and glycolysis of breast cancer cells by mediating the ubiquitination level of PFK-1. The USP35/PFK-1 axis offers novel insight for the treatment of breast cancer.
泛素特异性蛋白酶35（USP35）被发现参与多种肿瘤进展，但其在乳腺癌中的作用仍 largely未知。分别通过定量实时PCR和蛋白质免疫印迹法评估乳腺癌组织和细胞中USP35 mRNA和蛋白质的表达。随后，采用流式细胞术和5-乙炔基-2'-脱氧尿苷标记法评估乳腺癌细胞的凋亡和增殖。使用海马分析和各种试剂盒分析细胞糖酵解功能。此外，利用免疫共沉淀（Co-IP）和免疫沉淀试验验证USP35的去泛素化机制。最后，在裸鼠中建立皮下人异种移植肿瘤模型以验证USP35在体内的作用。通过检测临床样本和细胞系，我们发现USP35在乳腺癌中表达显著上调。进一步的功能研究表明，敲低USP35表达可抑制细胞增殖并促进凋亡。此外，与对照短发夹RNA（sh-Control）相比，敲低USP35可降低磷酸果糖激酶1（PFK-1）的表达，并与较低的细胞外酸化率和耗氧率相关。Co-IP试验确定PFK-1是USP35的直接去泛素化靶点。重要的是，我们证明PFK-1是USP35在体外和体内诱导细胞增殖和糖酵解的关键介质。本研究确定USP35通过介导PFK-1的泛素化水平来调节乳腺癌细胞的增殖和糖酵解。USP35/PFK-1轴为乳腺癌的治疗提供了新的见解。本研究确定USP35通过介导PFK-1的泛素化水平来调节乳腺癌细胞的增殖和糖酵解。USP35/PFK-1轴为乳腺癌的治疗提供了新的见解。
