School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia.
J Proteome Res. 2011 Mar 4;10(3):1030-42. doi: 10.1021/pr100803b. Epub 2011 Feb 4.
Cladribine (CdA) and fludarabine (FdAMP) are purine analogs that induce apoptosis in chronic lymphocytic leukemia and non-Hodgkin's lymphoma, but the mechanisms are undefined. The effects of CdA and fludarabine nucleoside (FdA) on the cytosolic, mitochondrial, and nuclear proteomes in human Raji lymphoma cells have been determined using two-dimensional fluorescence difference gel electrophoresis (DIGE) and mass spectrometry. Differentially abundant proteins have provided new insights into CdA- and FdA-induced apoptosis. Treatment with these purine analogs induced changes in proteins involved with intermediary metabolism, cell growth, signal transduction, protein metabolism, and regulation of nucleic acids. Differentially abundant mitochondrial 39S ribosomal protein L50, mTERF domain-containing protein 1, Chitinase-3 like 2 protein, and ubiquinone biosynthesis protein COQ9 have been identified in cells undergoing apoptosis. Up-regulation of several stress-associated proteins found in the endoplasmic reticulum (ER) including GRP78, ERp57, and ORP150 suggests that purine analog-induced apoptosis may result from ER stress and unfolded protein response. While mitochondria-dependent apoptosis has been associated with purine analog cytotoxicity, the likely involvement of the ER stress pathway in CdA- and FdA-induced apoptosis has been shown here for the first time.
克拉屈滨（CdA）和氟达拉滨（FdAMP）是嘌呤类似物，可诱导慢性淋巴细胞白血病和非霍奇金淋巴瘤细胞凋亡，但具体机制尚不清楚。本研究采用二维荧光差异凝胶电泳（DIGE）和质谱技术，检测了 CdA 和氟达拉滨核苷（FdA）对人 Raji 淋巴瘤细胞胞质溶胶、线粒体和核蛋白质组的影响。差异丰度蛋白为 CdA 和 FdA 诱导的细胞凋亡提供了新的见解。这些嘌呤类似物的处理诱导了与中间代谢、细胞生长、信号转导、蛋白质代谢和核酸调节相关的蛋白质发生变化。凋亡细胞中鉴定到差异丰度的线粒体 39S 核糖体蛋白 L50、mTERF 结构域包含蛋白 1、几丁质酶 3 样蛋白 2 和泛醌生物合成蛋白 COQ9。内质网（ER）中几种应激相关蛋白（包括 GRP78、ERp57 和 ORP150）的上调表明嘌呤类似物诱导的细胞凋亡可能是由于 ER 应激和未折叠蛋白反应所致。虽然线粒体依赖性细胞凋亡与嘌呤类似物的细胞毒性有关，但本研究首次表明 ER 应激途径可能参与了 CdA 和 FdA 诱导的细胞凋亡。
