Department of Biochemical Sciences, University of Florence, Viale G. Morgagni, 50, 50134, Florence, Italy.
J Biol Inorg Chem. 2010 May;15(4):573-82. doi: 10.1007/s00775-010-0624-3. Epub 2010 Jan 29.
We have recently shown that a group of structurally diverse gold compounds are highly cytotoxic toward a panel of 36 human tumor cell lines through a variety of biochemical mechanisms. A classic proteomic approach is exploited here to gain deeper insight into those mechanisms. This investigation is focused on Auoxo6, a novel binuclear gold(III) complex, and auranofin, a clinically established gold(I) antiarthritic drug. First, the 72-h cytotoxicity profiles of Auoxo6 and auranofin were determined against A2780 human ovarian carcinoma cells. Subsequently, protein extraction from gold-treated A2780 cells sensitive to cisplatin and 2D gel electrophoresis separation were carried out according to established procedures. Notably, both metallodrugs caused relatively modest changes in protein expression in comparison with controls as only 11 out of approximately 1,300 monitored spots showed appreciable quantitative changes. Very remarkably, six altered proteins were in common between the two treatments. Eight altered proteins were identified by mass spectrometry; among them was ezrin, a protein associated with the cytoskeleton and involved in apoptosis. Interestingly, two altered proteins, i.e., peroxiredoxins 1 and 6, are known to play crucial roles in the cell redox metabolism. Increased cleavage of heterogeneous ribonucleoprotein H was also evidenced, consistent with caspase 3 activation. Overall, the results of the present proteomic study point out that the mode of action of Auoxo6 is strictly related to that of auranofin, that the induced changes in protein expression are limited and selective, that both gold compounds trigger caspase 3 activation and apoptosis, and that a few affected proteins are primarily involved in cell redox homeostasis.
我们最近发现,一组结构多样的金化合物通过多种生化机制对 36 个人类肿瘤细胞系表现出高度细胞毒性。这里采用了经典的蛋白质组学方法来深入了解这些机制。本研究集中在 Auoxo6 上,一种新型双核金(III)配合物和临床应用的金(I)抗风湿药物。首先,确定了 Auoxo6 和金诺芬对 A2780 人卵巢癌细胞的 72 小时细胞毒性曲线。随后,根据既定程序,从对顺铂敏感的金处理的 A2780 细胞中提取蛋白质并进行 2D 凝胶电泳分离。值得注意的是,与对照相比,两种金属药物引起的蛋白质表达变化相对较小,只有大约 1300 个监测点中的 11 个显示出明显的定量变化。非常显著的是,两种处理之间有六个改变的蛋白质是共同的。通过质谱鉴定了 8 个改变的蛋白质;其中包括与细胞骨架相关并参与细胞凋亡的 ezrin。有趣的是,两种改变的蛋白质,即过氧化物酶 1 和 6,已知在细胞氧化还原代谢中起关键作用。还证明了异质核糖核蛋白 H 的增加切割,与 caspase 3 激活一致。总体而言,本蛋白质组学研究的结果表明,Auoxo6 的作用模式与金诺芬严格相关,诱导的蛋白质表达变化是有限和选择性的,两种金化合物均触发 caspase 3 激活和细胞凋亡,并且一些受影响的蛋白质主要参与细胞氧化还原稳态。