Panpan S I, Wei G E, Kaiming W U, Zhang Renquan
Department of Chest Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, 230031, Anhui, China.
Mol Cell Biochem. 2025 Apr;480(4):2633-2643. doi: 10.1007/s11010-024-05146-2. Epub 2024 Nov 4.
Non-small cell lung cancer (NSCLC) stands as the prevailing manifestation of lung cancer, with current therapeutic modalities linked to a dismal prognosis, necessitating further advancements. Hexokinase 2 (HK2), a critical enzyme positioned on the mitochondrial membrane, exerts control over diverse biological pathways, thereby regulating cancer. Nevertheless, the precise role and mechanism of HK2 in NSCLC remain inadequately elucidated, warranting comprehensive investigation. HK2 expression in NSCLC tissues and cell lines was detected through immunohistochemistry and western blot analysis. Concurrently, shRNA assays were applied to scrutinize the impact of HK2 on cell proliferation, apoptosis, migration, and invasion processes in NSCLC cell lines, utilizing CCK8, flow cytometry, wound-healing assay, and transwell techniques. The involvement of HK2 in mitochondrial dynamics was probed through western blot analysis, mitochondrial membrane potential assay, and assessment of ROS generation. Next, the functional role of HK2 was assessed by examining its influence on xenograft tumor growth in nude mice in vivo. Further research has demonstrated that HK2 played a role in NSCLC through its O-GlcNAcylation process. The results of the study revealed that HK2 O-GlcNAcylation promoted the proliferation, migration, and invasive characteristics of NSCLC cells, while alleviating mitochondrial damage, whereas O-GlcNAcylation inactivation yielded the opposite effect. Furthermore, in vivo experiments in nude mice illustrated that HK2 O-GlcNAcylation could stimulate tumor growth in NSCLC. These results suggested that HK2 may impact mitochondrial dynamics in NSCLC through its O-GlcNAcylation, thereby contributing to the progression of NSCLC.
非小细胞肺癌(NSCLC)是肺癌的主要表现形式,目前的治疗方式预后不佳,需要进一步改进。己糖激酶2(HK2)是一种位于线粒体膜上的关键酶,可控制多种生物途径,从而调节癌症。然而,HK2在NSCLC中的具体作用和机制仍未得到充分阐明,需要进行全面研究。通过免疫组织化学和蛋白质印迹分析检测NSCLC组织和细胞系中HK2的表达。同时,应用短发夹RNA(shRNA)检测来研究HK2对NSCLC细胞系中细胞增殖、凋亡、迁移和侵袭过程的影响,采用细胞计数试剂盒8(CCK8)、流式细胞术、伤口愈合试验和Transwell技术。通过蛋白质印迹分析、线粒体膜电位检测和活性氧(ROS)生成评估来探究HK2在线粒体动力学中的作用。接下来,通过检测HK2对裸鼠体内异种移植肿瘤生长的影响来评估其功能作用。进一步的研究表明,HK2通过其O-连接的N-乙酰葡糖胺(O-GlcNAc)糖基化过程在NSCLC中发挥作用。研究结果显示,HK2的O-GlcNAc糖基化促进了NSCLC细胞的增殖、迁移和侵袭特性,同时减轻了线粒体损伤,而O-GlcNAc糖基化失活则产生相反的效果。此外,裸鼠体内实验表明,HK2的O-GlcNAc糖基化可刺激NSCLC肿瘤生长。这些结果表明,HK2可能通过其O-GlcNAc糖基化影响NSCLC中的线粒体动力学,从而促进NSCLC的进展。