Jia Weiqiang, Zhang Yanling, Zhao Qiang, Gong Min, Cao Yang, Liu Jia, Luo Shuang
Cancer Prevention and Treatment Institute of Chengdu, Department of Neurosurgery, Chengdu Fifth People's Hospital (The Second Clinical Medical College, Affiliated Fifth People's Hospital of Chengdu University of Traditional Chinese Medicine), Chengdu 611137, China.
The First Clinical School of Guangzhou University of Chinese Medicine, Guangzhou 510000, China.
J Cancer. 2025 Jan 1;16(1):44-54. doi: 10.7150/jca.95266. eCollection 2025.
Glioblastoma (GBM), notorious for its poor prognosis, stands as a formidable challenge within the central nervous system tumor category, primarily due to its intricate pathology that encompasses stemness and the epithelial-mesenchymal transition (EMT). The ubiquity of S phase kinase-associated protein 2 (Skp2) overexpression in GBM, a protein implicated in both EMT and stemness traits, correlates with increased drug resistance, elevated tumor grades, and adverse outcomes. This investigation delves into the impact of Raddeanin A (RA), a triterpenoid compound extracted from Anemone raddeana Regel, on GBM, with a special focus on its influence over Skp2 expression levels. The study assessed RA's influence on GBM cell lines U87 and U251 via CCK-8 and colony formation assays to gauge cell proliferation, alongside Transwell assays for evaluating migration and invasion capabilities. mRNA expression was detected by RT-PCR. Protein expression alterations were examined through western blotting and immunofluorescence techniques. The therapeutic potential of RA was also evaluated using subcutaneous and intracranial xenograft model in mice, developed using U87 cells. The molecular docking experiment was performed to evaluate the binding of RA to Skp2. RA markedly curtailed the proliferation of U87 and U251 cells in a concentration-dependent manner, alongside diminishing sphere formation in glioblastoma stem-like cells (GSCs). A significant suppression of Skp2 expression was observed in both cell lines and GSCs following RA treatment. This reduction in Skp2 was associated with a decrease in stemness markers (Sox2, Nestin) and the inhibition of EMT markers (Vimentin, N-cadherin, Snail). Moreover, Skp2 overexpression was found to mitigate RA's suppressive effects on EMT and stemness, highlighting Skp2's crucial role in these processes. The experiments supported these findings, indicating that RA not only thwarted tumor growth but also substantially lowered the expression of Skp2, EMT markers, and stemness indicators. Additionally, molecular docking experiments demonstrated that RA exhibits a notable binding affinity to Skp2. This study elucidates RA's significant antitumor efficacy against GBM and by targeting pathways linked to stemness and EMT, chiefly via the downregulation of Skp2. These findings underscore RA's therapeutic promise in GBM management, offering insights into its mechanism of action and laying the groundwork for subsequent clinical investigations.
胶质母细胞瘤（GBM）因其预后不良而臭名昭著，是中枢神经系统肿瘤中的一个巨大挑战，主要是由于其复杂的病理过程，包括干性和上皮-间质转化（EMT）。S期激酶相关蛋白2（Skp2）在GBM中普遍过表达，该蛋白与EMT和干性特征均有关联，与耐药性增加、肿瘤分级升高及不良预后相关。本研究探讨了从毛茛科植物红升麻中提取的三萜类化合物升麻素A（RA）对GBM的影响，特别关注其对Skp2表达水平的影响。该研究通过CCK-8和集落形成试验评估RA对GBM细胞系U87和U251的影响，以衡量细胞增殖，同时通过Transwell试验评估迁移和侵袭能力。通过RT-PCR检测mRNA表达。通过蛋白质印迹和免疫荧光技术检测蛋白质表达变化。还使用U87细胞建立的小鼠皮下和颅内异种移植模型评估了RA的治疗潜力。进行分子对接实验以评估RA与Skp2的结合情况。RA以浓度依赖性方式显著抑制U87和U251细胞的增殖，并减少胶质母细胞瘤干细胞样细胞（GSCs）中的球状体形成。RA处理后，在这两种细胞系和GSCs中均观察到Skp2表达的显著抑制。Skp2的这种减少与干性标志物（Sox2、Nestin）的减少以及EMT标志物（波形蛋白、N-钙黏蛋白、Snail）的抑制有关。此外，发现Skp2过表达可减轻RA对EMT和干性的抑制作用，突出了Skp2在这些过程中的关键作用。体内实验支持了这些发现，表明RA不仅阻碍肿瘤生长，还显著降低了Skp2、EMT标志物和干性指标的表达。此外，分子对接实验表明RA对Skp2具有显著的结合亲和力。本研究阐明了RA对GBM具有显著的抗肿瘤疗效，并且通过靶向与干性和EMT相关的途径，主要是通过下调Skp2来实现。这些发现强调了RA在GBM治疗中的前景，为其作用机制提供了见解，并为后续的临床研究奠定了基础。
