Jiang Kainian, Yang Wei, Huang Jie, Tan Xiaolong, Liu Yan, Tu Saiya, Luo Jian
Department of Geriatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China.
Department of Hepatobiliary and Pancreatic Surgery, The Third People's Hospital of Hubei Province, Wuhan, Hubei 430030, P.R. China.
Int J Mol Med. 2025 Sep;56(3). doi: 10.3892/ijmm.2025.5582. Epub 2025 Jul 19.
SUMOylation is a critical post‑translational modification, serving as a key role in nucleocytoplasmic translocation, transcriptional cofactor stabilization and modulation of chromatin remodeling factors, which are associated with oncogenesis, tumor progression and chemotherapy resistance in various types of cancer. SUMOylation was performed by small ubiquitin‑like modifier (SUMO), a kind of small ubiquitin‑like modifier, which was attached or removed from the substrates. The excessive export of nuclear p27kip1 induced by SUMOylation is associated with cell proliferation and chemotherapy resistance in cholangiocarcinoma (CCA). However, the exact underlying mechanism remains currently unknown. The present study investigated SUMO specific peptidase 1 (SENP1), which is known to participate in SUMOylation by activating nuclear SUMO1 precursors and deSUMOylating cytoplasmic substrates. SENP1 exhibited increased expression levels in CCA specimens compared with that in adjacent non‑cancerous tissues, as confirmed by bioinformatics analysis and immunohistochemical assays. A significant correlation between SENP1 and p27kip1 expression levels was observed. SENP1 overexpression significantly increased cytoplasmic p27kip1 expression levels, thereby promoting CCA cell proliferation, accelerating the G1‑S cell cycle transition and reducing chemical sensitivity through increasing overall SUMOylation of p27kip1, as confirmed via western blotting, immunofluorescence, flow cytometry, Cell Counting Kit‑8, 5‑ethynyl‑2'‑deoxyuridine incorporation and SUMOylation tests. By contrast, SENP1 knockdown demonstrated the opposite results. Subsequently, the use of ML‑792, COH000 and leptomycin B treatments, and the mutant variant SENP1‑C603A demonstrated that SENP1 regulates the functionality of p27kip1 through nuclear SUMOylation rather than cytoplasmic deSUMOylation. The involvement of SENP1 represents a pivotal role in governing the nucleocytoplasmic shuttling of p27kip1. SENP1 knockdown could effectively impede CCA cell proliferation and enhance the chemosensitivity of cis‑platinum by modulating the nuclear export of p27kip1 through SUMOylation, thus offering a potential therapeutic approach for CCA in the future.
小泛素样修饰（SUMO）化是一种关键的翻译后修饰，在核质转运、转录辅因子稳定以及染色质重塑因子的调节中发挥关键作用，这些过程与多种癌症的肿瘤发生、肿瘤进展及化疗耐药相关。SUMO化由小泛素样修饰物（SUMO）进行，SUMO是一种小泛素样修饰物，可与底物结合或从底物上移除。SUMO化诱导的核p27kip1过度输出与胆管癌（CCA）的细胞增殖及化疗耐药相关。然而，确切的潜在机制目前仍不清楚。本研究调查了SUMO特异性蛋白酶1（SENP1），已知其通过激活核SUMO1前体并使细胞质底物去SUMO化来参与SUMO化过程。生物信息学分析和免疫组织化学检测证实，与相邻非癌组织相比，SENP1在CCA标本中的表达水平升高。观察到SENP1与p27kip1表达水平之间存在显著相关性。通过蛋白质免疫印迹、免疫荧光、流式细胞术、细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷掺入及SUMO化检测证实，SENP1过表达显著增加细胞质p27kip1表达水平，从而促进CCA细胞增殖，加速G1-S细胞周期转变，并通过增加p27kip1的整体SUMO化降低化学敏感性。相比之下，敲低SENP1则产生相反的结果。随后，使用ML-792、COH000和雷帕霉素B处理以及突变体变体SENP1-C603A表明，SENP1通过核SUMO化而非细胞质去SUMO化来调节p27kip1的功能。SENP1的参与在控制p27kip1的核质穿梭中起关键作用。敲低SENP1可通过SUMO化调节p27kip1的核输出，有效抑制CCA细胞增殖并增强顺铂的化学敏感性，从而为未来CCA提供一种潜在的治疗方法。
