Bada Lucía, Butt Hussain Shakeel, Quezada Elías, Picos Aitor, Wangensteen Helle, Inngjerdingen Kari Tvete, Gil-Longo José, Viña Dolores
Group of Pharmacology of Chronic Diseases (CD Pharma), Molecular Medicine and Chronic Diseases Research Centre (CIMUS), Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
Department of Pharmacology, Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
Molecules. 2025 Feb 19;30(4):972. doi: 10.3390/molecules30040972.
The plant kingdom serves as a valuable resource for cancer drug development. This study explored the antitumor activity of different sub-fractions (hexane, dichloromethane and methanol) of (gorse) methanol extract in glioblastoma (U-87MG and U-373MG) and neuroblastoma (SH-SY5Y) cell lines, along with their phytochemical profiles. Cytotoxicity was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays, and cell cycle arrest and apoptosis were assessed through flow cytometry and by measuring reactive oxygen species (ROS) and protein expression levels. D7 and D8 dichloromethane sub-fractions significantly reduced cell viability, triggered early apoptosis in SH-SY5Y and U-87MG cells and specifically increased ROS levels in U-87MG cells. Western blot analyses showed that D7 increased p53, caspase-3, caspase-8 and γH2AX expression in SH-SY5Y and U-87MG cells, while D8 specifically elevated p53 in SH-SY5Y cells and caspase-3 in both cell lines. In U-373 cells, D7 and D8 markedly reduced cell viability, with D8 inducing necrosis. Morphological changes indicative of apoptosis were also observed in all cell lines. Bioinformatic analysis of UHPLC-MS and GC-MS data tentatively identified 20 metabolites in D7 and 15 in D8, primarily flavonoids. HPLC-DAD confirmed isoprunetin and genistein as the most abundant in D7 and D8, respectively, both isolated and identified by NMR spectroscopy. Most of the flavonoids identified have been reported as antitumor agents, suggesting that these compounds may be responsible for the observed pharmacological activity.
植物王国是癌症药物开发的宝贵资源。本研究探讨了金雀花甲醇提取物的不同亚组分(己烷、二氯甲烷和甲醇)对胶质母细胞瘤(U-87MG和U-373MG)和神经母细胞瘤(SH-SY5Y)细胞系的抗肿瘤活性及其植物化学特征。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)和乳酸脱氢酶(LDH)测定法评估细胞毒性,并通过流式细胞术以及测量活性氧(ROS)和蛋白质表达水平来评估细胞周期阻滞和凋亡。二氯甲烷亚组分D7和D8显著降低细胞活力,在SH-SY5Y和U-87MG细胞中引发早期凋亡,并特异性增加U-87MG细胞中的ROS水平。蛋白质印迹分析表明,D7增加了SH-SY5Y和U-87MG细胞中p53、半胱天冬酶-3、半胱天冬酶-8和γH2AX的表达,而D8特异性提高了SH-SY5Y细胞中的p53以及两种细胞系中的半胱天冬酶-3。在U-373细胞中,D7和D8显著降低细胞活力,D8诱导坏死。在所有细胞系中也观察到了指示凋亡的形态学变化。对超高效液相色谱-质谱(UHPLC-MS)和气相色谱-质谱(GC-MS)数据的生物信息学分析初步鉴定出D7中有20种代谢物,D8中有15种,主要为黄酮类化合物。高效液相色谱-二极管阵列检测(HPLC-DAD)证实异普林汀和染料木黄酮分别是D7和D8中含量最高的成分,二者均通过核磁共振光谱法分离和鉴定。已报道所鉴定出的大多数黄酮类化合物为抗肿瘤剂,这表明这些化合物可能是观察到的药理活性的原因。