Chen Xiaoqing, Su Weijie, Chen Jiewen, Ouyang Peng, Gong Jin
Department of Breast Medicine, The affiliated Foshan Women and Children Hospital, Guangdong Medical University, Foshan, 528000, China.
Department of Gastrointestinal Surgery, The First Affiliated Hospital of Jinan University, 613 West of Huangpu Avenue, Guangzhou, 510630, China.
Naunyn Schmiedebergs Arch Pharmacol. 2024 Dec 27. doi: 10.1007/s00210-024-03723-2.
E3 ubiquitin ligases have the potential to modulate key oncogenic pathways. RING finger protein 123 (RNF123), as an E3 ubiquitin ligase, has been functioned as a tumor suppressor. This study was designed to explore the role of RNF123 in breast cancer. Immunohistochemistry was applied to examine protein expression in breast cancer tissues. Western blot and Quantitative Real-time PCR were performed to gauge protein and mRNA levels. Lentivirus transduction was used to overexpress or silence genes of interest. Cell Counting Kit-8, flow cytometry, and colony formation assays were used to assess cell viability, cell cycle, and colony formation. Extracellular acidification rate, lactic acid and adenosine triphosphate were used for glycolysis assay. Co-immunoprecipitation (Co-IP) and ubiquitination analysis were used to explore the interaction between RNF123 and 6-Phosphofructo-2-kinase (PFKP). In vivo experiments were performed with xenograft tumor models. RNF123 was downregulated in tumor tissues and cells, overexpression of which significantly decreased the viability and colony-forming ability of tumor cells, suppressed the progression of the cell cycle and glycolytic activity, and suppressed tumor growth in vivo. Co-IP and ubiquitination analysis revealed that there was an interaction between RNF123 and PFKP, and RNF123 could induce ubiquitination of PFKP. PFKP could reverse the effects of RNF123 on tumor cells. RNF123 inhibited cell viability, cell cycle and colony formation of breast cancer cells by inhibiting glycolysis via ubiquitination of PFKP.
E3泛素连接酶有调节关键致癌途径的潜力。环指蛋白123（RNF123）作为一种E3泛素连接酶，发挥着肿瘤抑制因子的作用。本研究旨在探讨RNF123在乳腺癌中的作用。应用免疫组织化学检测乳腺癌组织中的蛋白表达。进行蛋白质免疫印迹法和定量实时聚合酶链反应以测定蛋白质和mRNA水平。使用慢病毒转导来过表达或沉默目的基因。使用细胞计数试剂盒-8、流式细胞术和集落形成试验来评估细胞活力、细胞周期和集落形成。细胞外酸化率、乳酸和三磷酸腺苷用于糖酵解测定。免疫共沉淀（Co-IP）和泛素化分析用于探索RNF123与6-磷酸果糖-2-激酶（PFKP）之间的相互作用。在异种移植肿瘤模型上进行体内实验。RNF123在肿瘤组织和细胞中表达下调，其过表达显著降低肿瘤细胞的活力和集落形成能力，抑制细胞周期进程和糖酵解活性，并在体内抑制肿瘤生长。免疫共沉淀和泛素化分析显示RNF123与PFKP之间存在相互作用，且RNF123可诱导PFKP的泛素化。PFKP可逆转RNF123对肿瘤细胞的作用。RNF123通过使PFKP泛素化抑制糖酵解，从而抑制乳腺癌细胞的活力、细胞周期和集落形成。
