Wang Xinxing, Ma Mingze, Shao Shuai, Xu Xianwen, Qin Chuan, Gao Ruxin, Zhang Zhenhai
Department of Hepatobiliary Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, 250021, China.
Departments of Infectious Diseases, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, 250021, China.
Cancer Cell Int. 2025 Feb 7;25(1):37. doi: 10.1186/s12935-024-03583-z.
TWIST1 is known to promote glycolysis and contribute to pancreatic cancer development; however, its underlying mechanisms remain poorly understood. This study aims to elucidate the molecular mechanisms by which TWIST1 influences aerobic glycolysis in pancreatic ductal adenocarcinoma (PDAC).
The expression levels of TWIST1, MMP9, MT1-MMP, and FDX1 in clinical tissues and cancer cell lines were assessed using quantitative reverse transcription PCR (QRT-PCR). Cell treatments with Elesclomol-Cu and 2-deoxyglucose (2DG) were conducted. Immunofluorescence staining and immunoprecipitation analyses were performed to investigate the binding relationship between TWIST1 and HK2. Colony formation and Transwell assays were utilized to evaluate the effects of TWIST1 on cell proliferation, migration, and invasion. Western blotting was employed to detect proteins related to cuproptosis and apoptosis, while ubiquitination assays assessed TWIST1's regulation of HK2 ubiquitination.
TWIST1 expression was significantly elevated in PDAC tissues, and over-expression of TWIST1 in PDAC cells enhanced colony formation and cell proliferation. Notably, HK2 levels were markedly higher in pancreatic cancer tissues compared to adjacent normal tissues. TWIST1 was found to directly bind and interact with HK2, showing co-localization in the cytoplasm of PDAC cells. Furthermore, TWIST1 was shown to stabilize HK2 by inhibiting its ubiquitin-mediated degradation. Knockdown of TWIST1 or HK2 enhanced the inhibitory effects of 2DG on cell migration and invasion. Treatment with Elesclomol-Cu and 2DG significantly reduced the expression of the cuproptosis-related factor FDX1 with no impact on other cell death factors.
This study demonstrates that TWIST1 regulates the ubiquitination and degradation of HK2, thereby promoting glycolysis-induced cuproptosis and facilitating pancreatic cancer invasion and metastasis. Understanding the underlying mechanisms of PDAC, including the regulation of key proteins such as HK2 by TWIST1, is crucial for developing more effective treatment strategies. Findings highlight the importance of targeting these molecular pathways, which could lead to improved diagnostic and therapeutic approaches, ultimately enhancing patient outcomes and prognosis.
已知TWIST1可促进糖酵解并推动胰腺癌发展；然而，其潜在机制仍知之甚少。本研究旨在阐明TWIST1影响胰腺导管腺癌（PDAC）有氧糖酵解的分子机制。
使用定量逆转录PCR（QRT-PCR）评估临床组织和癌细胞系中TWIST1、MMP9、MT1-MMP和FDX1的表达水平。用依列莫莫尔-铜（Elesclomol-Cu）和2-脱氧葡萄糖（2DG）进行细胞处理。进行免疫荧光染色和免疫沉淀分析以研究TWIST1与HK2之间的结合关系。利用集落形成和Transwell实验评估TWIST1对细胞增殖、迁移和侵袭的影响。采用蛋白质印迹法检测与铜死亡和凋亡相关的蛋白质，同时通过泛素化实验评估TWIST1对HK2泛素化的调节作用。
TWIST1在PDAC组织中的表达显著升高，且PDAC细胞中TWIST1的过表达增强了集落形成和细胞增殖。值得注意的是，与相邻正常组织相比，胰腺癌组织中HK2水平明显更高。发现TWIST1直接与HK2结合并相互作用，在PDAC细胞的细胞质中共定位。此外，TWIST1通过抑制HK2泛素介导的降解来稳定HK2。敲低TWIST1或HK2增强了2DG对细胞迁移和侵袭的抑制作用。用依列莫莫尔-铜和2DG处理显著降低了铜死亡相关因子FDX1的表达，对其他细胞死亡因子无影响。
本研究表明，TWIST1调节HK2的泛素化和降解，从而促进糖酵解诱导的铜死亡并促进胰腺癌的侵袭和转移。了解PDAC的潜在机制，包括TWIST1对HK2等关键蛋白的调节，对于制定更有效的治疗策略至关重要。研究结果突出了靶向这些分子途径的重要性，这可能导致改进的诊断和治疗方法，最终改善患者的结局和预后。
