Güçlü Ebru, Ayan İlknur Çınar, Çetinkaya Sümeyra, Dursun Hatice Gül, Vural Hasibe
Department of Basic Science and Health, Hemp Research Institute, Yozgat Bozok University, Yozgat, Turkey.
Department of Medical Biology, Meram Faculty of Medicine, Necmettin Erbakan University, Konya, Turkey.
J Appl Toxicol. 2024 Nov;44(11):1714-1724. doi: 10.1002/jat.4671. Epub 2024 Jul 14.
The aim of this study was to evaluate the anticancer effects of piceatannol, a natural stilbenoid, on human neuroblastoma cells. In order to accomplish this goal, we performed various cellular assays, including the XTT cell proliferation assay for cell viability, colony formation assay for colony formation capacity, FITC Annexin V and cell death detection kit for apoptosis, matrigel invasion assay for invasion capacity, intracellular reactive oxygen species (ROS) red dye for intracellular ROS levels, TMRM staining method for mitochondrial membrane potential (MMP), and the CYTO-ID autophagy detection kit for autophagy. Furthermore, we analyzed the expression levels of genes associated with apoptosis and autophagy using RT-qPCR. Based on our findings, piceatannol exhibited cytotoxic effects on neuroblastoma cells. Besides, treatment with piceatannol at both 50 and 100 μM concentrations for 72 h decreased colony formation, induced apoptosis and autophagy, inhibited cell invasion, decreased MMP, and increased ROS levels in SH-SY5Y cells. In addition, we observed significant upregulation in the expression levels of CASP8, BECLIN, ATG5, ATG7, and MAPILC3A genes between the two doses. These results suggest that piceatannol enhances autophagic activity and induces caspase-dependent apoptosis, indicating its potential as a therapeutic agent against neuroblastoma cells.
本研究的目的是评估天然芪类化合物白皮杉醇对人神经母细胞瘤细胞的抗癌作用。为实现这一目标，我们进行了各种细胞试验，包括用于检测细胞活力的XTT细胞增殖试验、用于检测集落形成能力的集落形成试验、用于检测凋亡的FITC膜联蛋白V和细胞死亡检测试剂盒、用于检测侵袭能力的基质胶侵袭试验、用于检测细胞内活性氧（ROS）水平的细胞内ROS红色染料、用于检测线粒体膜电位（MMP）的TMRM染色法以及用于检测自噬的CYTO-ID自噬检测试剂盒。此外，我们使用RT-qPCR分析了与凋亡和自噬相关的基因表达水平。基于我们的研究结果，白皮杉醇对神经母细胞瘤细胞具有细胞毒性作用。此外，在SH-SY5Y细胞中，用50和100μM浓度的白皮杉醇处理72小时可减少集落形成、诱导凋亡和自噬、抑制细胞侵袭、降低MMP并增加ROS水平。此外，我们观察到两剂量之间CASP8、BECLIN、ATG5、ATG7和MAPILC3A基因的表达水平显著上调。这些结果表明，白皮杉醇可增强自噬活性并诱导半胱天冬酶依赖性凋亡，表明其作为抗神经母细胞瘤细胞治疗剂的潜力。
