Xu De-Lai, Fan Kai, Zhang Hua, Tang Liu-Xing, Wang Yang, Xiang Zhen, Shi Ai-Ming, Qu Yu-Chen, Su Cun-Jin, Pan Jie
Department of Pharmacy, the Second Affiliated Hospital of Soochow University, Suzhou, China.
Transl Pediatr. 2022 Aug;11(8):1346-1361. doi: 10.21037/tp-22-331.
In childhood, metastatic neuroblastoma (NB) is the most common extracranial solid tumor, but there are no appropriate drugs for its treatment. Dihydroartemisinin (DHA), a drug for malaria treatment, has therapeutic potential in several cancers; however, its mechanisms remain unclear. This study aimed to investigate the anti-proliferation effect of DHA on SH-SY5Y cells and to explore its mechanism .
We used 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to measure the half-maximal inhibitory concentration (IC50) of DHA; western blot was used to determine protein levels; propidium iodide (PI) staining was used to determine apoptotic cells; JC-1 staining to measure mitochondrial membrane potential; and dichloro-dihydro-fluorescein diacetate (DCFH-DA) staining was used to determine reactive oxygen species (ROS). Metabonomic analysis was performed by using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)-based untargeted metabolomics. Multivariate statistical analysis was performed to screen potential metabolites associated with DHA treatment in SH-SY5Y cells.
It was shown that DHA inhibited SH-SY5Y cell proliferation and increased poly (ADP-ribose) polymerase (PARP-1) and caspase 3 in a dose-dependent manner. In Further, DHA promoted ROS generation and γH2AX expression. In addition, a total of 125 proposed metabolites in SH-SY5Y cells and 45 vital metabolic pathways were identified through UHPLC-MS/MS-based untargeted metabolomic analysis.
These data suggest that DHA could regulate taurine, linoleic acid, phenylalanine metabolism, and tryptophan metabolism, which are involved in the anti-proliferation effect of DHA in SH-SY5Y cells.
在儿童时期，转移性神经母细胞瘤（NB）是最常见的颅外实体瘤，但尚无合适的治疗药物。双氢青蒿素（DHA）是一种治疗疟疾的药物，在多种癌症中具有治疗潜力；然而，其作用机制尚不清楚。本研究旨在探讨DHA对SH-SY5Y细胞的抗增殖作用及其机制。
我们采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法测定DHA的半数抑制浓度（IC50）；采用蛋白质免疫印迹法测定蛋白质水平；采用碘化丙啶（PI）染色法测定凋亡细胞；采用JC-1染色法测定线粒体膜电位；采用二氯二氢荧光素二乙酸酯（DCFH-DA）染色法测定活性氧（ROS）。通过基于超高效液相色谱-串联质谱（UHPLC-MS/MS）的非靶向代谢组学进行代谢组学分析。进行多变量统计分析以筛选与DHA处理SH-SY5Y细胞相关的潜在代谢物。
结果表明，DHA以剂量依赖性方式抑制SH-SY5Y细胞增殖，并增加聚（ADP-核糖）聚合酶（PARP-1）和半胱天冬酶3的表达。此外，DHA促进ROS生成和γH2AX表达。此外，通过基于UHPLC-MS/MS的非靶向代谢组学分析，在SH-SY5Y细胞中总共鉴定出125种潜在代谢物和45条重要代谢途径。
这些数据表明，DHA可以调节牛磺酸、亚油酸、苯丙氨酸代谢和色氨酸代谢，这些代谢过程参与了DHA对SH-SY5Y细胞的抗增殖作用。
