Carlisi Daniela, D'Anneo Antonella, Martinez Roberta, Emanuele Sonia, Buttitta Giuseppina, Di Fiore Riccardo, Vento Renza, Tesoriere Giovanni, Lauricella Marianna
Laboratory of Biochemistry, Department of Experimental Biomedicine and Clinical Neurosciences, Polyclinic, University of Palermo, 90127 Palermo, Italy.
Laboratory of Biochemistry, Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, Polyclinic, University of Palermo, 90127 Palermo, Italy.
Oncol Rep. 2014 Jul;32(1):167-72. doi: 10.3892/or.2014.3212. Epub 2014 May 23.
It has been shown that the sesquiterpene lactone parthenolide lowers the viability of MDA-MB-231 breast cancer cells, in correlation with oxidative stress. The present report examined the different radical species produced during parthenolide treatment and their possible role in the toxicity caused by the drug. Time course experiments showed that in the first phase of treatment (0-8 h), and in particular in the first 3 h, parthenolide induced dichlorofluorescein (DCF) signal in a large percentage of cells, while dihydroethidium (DHE) signal was not stimulated. Since the effect on DCF signal was suppressed by apocynin and diphenyleneiodonium (DPI), two inhibitors of NADPH oxidase (NOX), we suggest that parthenolide rapidly stimulated NOX activity with production of superoxide anion (O2•-), which was converted by superoxide dismutase 1 (SOD1) into hydrogen peroxide (H2O2). In the second phase of treatment (8-16 h), parthenolide increased the number of positive cells to DHE signal. Since this event was not prevented by apocynin and DPI and was associated with positivity of cells to MitoSox Red, a fluorochrome used to detect mitochondrial production of O2•-, we suggest that parthenolide induced production of O2•- at the mitochondrial level independently by NOX activity in the second phase of treatment. Finally, in this phase, most cells became positive to hydroxyphenyl fluorescein (HPF) signal, a fluorescent probe to detect highly reactive oxygen species (hROS), such as hydroxyl radical and peroxynitrite. Therefore, parthenolide between 8-16 h of treatment induced generation of O2•- and hROS, in close correlation with a marked reduction in cell viability.
已表明倍半萜内酯小白菊内酯可降低MDA-MB-231乳腺癌细胞的活力，这与氧化应激相关。本报告研究了小白菊内酯处理过程中产生的不同自由基种类及其在该药物所致毒性中的可能作用。时间进程实验表明，在处理的第一阶段（0 - 8小时），尤其是最初3小时，小白菊内酯在大部分细胞中诱导二氯荧光素（DCF）信号，而二氢乙锭（DHE）信号未被刺激。由于NADPH氧化酶（NOX）的两种抑制剂夹竹桃麻素和二亚苯基碘鎓（DPI）抑制了对DCF信号的影响，我们认为小白菊内酯迅速刺激NOX活性产生超氧阴离子（O2•-），其被超氧化物歧化酶1（SOD1）转化为过氧化氢（H2O2）。在处理的第二阶段（8 - 16小时），小白菊内酯增加了DHE信号阳性细胞的数量。由于这一事件未被夹竹桃麻素和DPI阻止，且与细胞对MitoSox Red（一种用于检测线粒体产生O2•-的荧光染料）呈阳性相关，我们认为在处理的第二阶段小白菊内酯在线粒体水平独立于NOX活性诱导O2•-的产生。最后，在这个阶段，大多数细胞对羟基苯基荧光素（HPF）信号呈阳性，HPF是一种用于检测高活性氧物种（hROS）如羟基自由基和过氧亚硝酸盐的荧光探针。因此，在处理8 - 16小时期间，小白菊内酯诱导O2•-和hROS的产生，这与细胞活力的显著降低密切相关。
