A proteome analysis of the arsenite response in cultured lung cells:: evidence for in vitro oxidative stress-induced apoptosis
A proteome analysis of the arsenite response in cultured lung cells:: evidence for in vitro oxidative stress-induced apoptosis
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DOI：
10.1042/bj20040224
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发表时间：
2004-09-01
影响因子：
4.1
通讯作者：
Chiu, JF
中科院分区：
文献类型：
--
作者：
Lau, ATY;He, QY;Chiu, JF
亚砷酸盐被认为是一种能够诱导细胞死亡的化疗药物。然而，在分子水平上的细胞反应还没有得到广泛的研究。在本研究中，我们首次对亚砷酸盐处理的大鼠肺上皮细胞(LECs)进行了蛋白质组学分析，目的是鉴定在砷诱导的细胞凋亡过程中可能表达的保护细胞的防御蛋白。对晶状体上皮细胞和40微米亚砷酸盐处理的晶状体上皮细胞进行比较蛋白质组分析，以确定其蛋白质表达谱的整体变化。用双向电泳法分离出1000多个蛋白质斑点，并用银染显影。通过基质辅助激光解吸电离飞行时间质谱仪和数据库检索鉴定了7种蛋白质的表达水平。上调的蛋白质主要是热休克蛋白和抗氧化应激蛋白，包括HSP70、醛糖还原酶、血红素加氧酶-1、HSP27、铁蛋白轻链和α-晶体蛋白。糖酵解酶甘油醛-3-磷酸脱氢酶表达下调。在亚砷酸盐伤害前用硫醇抗氧化剂谷胱甘肽或N-乙酰半胱氨酸预处理有效地消除了这些防御蛋白的诱导并维持了细胞的存活，而抗氧化剂只有在亚砷酸盐之后加入到细胞中才有保护作用。综上所述，我们的结果表明，高水平的亚砷酸盐会导致氧化应激诱导的细胞凋亡。
Arsenite is well documented as a chemotherapeutic agent capable of inducing cell death. However, the cellular response at the molecular level has not been studied extensively. In the present study, we provide for the first time a proteomic analysis of rat LECs (lung epithelial cells) treated with arsenite, with the aim of identifying defence proteins, probably expressed to protect the cells during the course of arsenic-induced apoptosis. Comparative proteome analysis was conducted on LECs and LECs treated with 40 muM arsenite to identify global changes in their protein expression profiles. Over 1000 protein spots were separated by two-dimensional electrophoresis and visualized by silver staining. Seven proteins changed expression levels significantly and were identified by matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry and database searching. The proteins up-regulated were mostly HSPs (heat-shock proteins) and antioxidative stress proteins, including HSP70, aldose reductase, haem oxygenase-1, HSP27, ferritin light chain and alphaB-crystallin. The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase was down-regulated. Pretreatment with the thiol antioxidants glutathione or N-acetylcysteine before arsenite insult effectively abrogated the induction of these defence proteins and sustained cell viability, whereas antioxidants were protective only at earlier time points if they were added to cells after arsenite. Taken together, our results demonstrate that high levels of arsenite cause oxidative stress-induced apoptosis.