Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Avenida Professor Lineu Prestes, 1374, 05508-000 São Paulo, SP, Brazil.
Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Avenida Professor Lineu Prestes, 1374, 05508-000 São Paulo, SP, Brazil.
Free Radic Biol Med. 2016 Jan;90:91-100. doi: 10.1016/j.freeradbiomed.2015.11.008. Epub 2015 Nov 11.
Chloroquine (CQ), a quinolone derivative widely used to treat and prevent malaria, has been shown to exert a potent adjuvant effect when combined with conventional glioblastoma therapy. Despite inducing lysosome destabilization and activating p53 in human glioma cells, the mechanisms underlying cell death induced by this drug are poorly understood. Here, we analyzed in a time- and dose-dependent manner, the effects of CQ upon mitochondria integrity, autophagy regulation and redox processes in four human glioma cell lines that differ in their resistance to this drug. NAC-containing media protected cells against CQ-induced loss of mitochondrial membrane potential (MMP), autophagic vacuoles (LC3II) accumulation and loss of cell viability induced by CQ. However, we noticed that part of this protection was due to media acidification in NAC preparations, alerting for problems in experimental procedures using NAC. The results indicate that although CQ induces accumulation of LC3II, mitochondria, and oxidative stress, neither of these events is clearly correlated to cell death induced by this drug. The only event elicited in all cell lines at equitoxic doses of CQ was the loss of MMP, indicating that mitochondrial stability is important for cells resistance to this drug. Finally, the data indicate that higher steady-state MMP values can predict cell resistance to CQ treatment.
氯喹(CQ)是一种广泛用于治疗和预防疟疾的喹啉衍生物,当与常规胶质母细胞瘤治疗联合使用时,已被证明具有强大的佐剂作用。尽管这种药物在人类神经胶质瘤细胞中诱导溶酶体不稳定并激活 p53,但诱导细胞死亡的机制仍知之甚少。在这里,我们在时间和剂量依赖性的方式下分析了 CQ 对四种对这种药物具有不同耐药性的人神经胶质瘤细胞系中线粒体完整性、自噬调节和氧化还原过程的影响。含 NAC 的培养基可防止 CQ 诱导的线粒体膜电位(MMP)丧失、自噬小泡(LC3II)积累和 CQ 诱导的细胞活力丧失。然而,我们注意到,这种保护的一部分是由于 NAC 制剂中的培养基酸化引起的,这就提醒我们在使用 NAC 的实验过程中存在问题。结果表明,尽管 CQ 诱导 LC3II、线粒体和氧化应激的积累,但这些事件都与该药物诱导的细胞死亡没有明显相关性。在 CQ 的等毒性剂量下,所有细胞系中唯一引发的事件是 MMP 的丧失,这表明线粒体稳定性对于细胞对该药物的耐药性很重要。最后,数据表明,更高的稳定态 MMP 值可以预测细胞对 CQ 治疗的耐药性。