Comparative proteomics analysis of sodium selenite-induced apoptosis in human prostate cancer cells
Comparative proteomics analysis of sodium selenite-induced apoptosis in human prostate cancer cells
复制标题
亚硒酸钠诱导人前列腺癌细胞凋亡的比较蛋白质组学分析
DOI：
10.1039/c3mt00002h
复制
发表时间：
2013-01-01
期刊：
影响因子：
3.4
通讯作者：
Ni, Jiazuan
中科院分区：
文献类型：
--
作者：
Chen, Ping;Wang, Lixiang;Ni, Jiazuan
硒是人体健康必需的重要微量矿物质。临床试验表明，硒对晚期或侵袭性前列腺癌具有潜在的抑制作用。然而，其作用机制仍不清楚。本研究利用蛋白质组学研究了亚硒酸钠在人前列腺癌 PC-3 细胞中的作用机制。 CCK-8法用于检测细胞活力和抑制率。流式细胞术通过膜联蛋白V-FITC和碘化丙啶双染检测细胞凋亡。亚硒酸盐抑制 PC-3 细胞的生长，导致它们表现出典型的细胞凋亡的形态变化。细胞凋亡率也增加。蛋白质组学鉴定了暴露于亚硒酸盐的 PC-3 细胞中多种差异表达的蛋白质。通过 MALDI-TOF 质谱法鉴定了 18 个蛋白质点。这些蛋白质被分为参与氧化还原平衡、蛋白质降解和细胞能量代谢的蛋白质。选择三种不同表达的蛋白（SOD1、Stathmin 和 Erp29）以及几种凋亡相关蛋白进行蛋白质印迹验证。 Western blot 分析表明，亚硒酸盐诱导的细胞凋亡伴随着 caspase-8 的激活和 PARP 的特异性蛋白水解切割。这导致促凋亡蛋白 Bax 增加，并导致抗凋亡蛋白 Bcl-2 和缺氧诱导因子 1 α 减少。 ROS 生成增加和线粒体膜电位降低与比较蛋白质组学鉴定的抗氧化蛋白表达减少一致。因此，我们认为亚硒酸钠主要通过线粒体途径诱导 PC-3 细胞凋亡，但也通过 ER 应激和 HIF-1 α 介导的途径。
Selenium is an important trace mineral necessary for human health. Clinical trials have shown potential inhibitory effects of selenium in advanced or aggressive prostate cancer. However, its mechanism of action remains unclear. This study investigated the mechanism of action of sodium selenite in human prostate cancer PC-3 cells using proteomics. CCK-8 assays were used to detect cell viability and the inhibitory rate. Cell apoptosis was detected by annexin V-FITC and propidium iodide double staining using flow cytometry. Selenite inhibited the growth of PC-3 cells causing them to display morphological changes typical of apoptosis. The rate of cell apoptosis also increased. Proteomics identified a variety of differentially expressed proteins in PC-3 cells exposed to selenite. Eighteen protein spots were identified by MALDI-TOF mass spectrometry. These proteins were separated into those involved in redox balance, protein degradation and cellular energy metabolism. Three differently expressed proteins (SOD1, Stathmin and Erp29) were chosen for Western blot verification, together with several apoptosis-related proteins. Western blot analyses showed that selenite-induced apoptosis was accompanied by activation of caspase-8 and specific proteolytic cleavage of PARP. This led to an increase in the pro-apoptotic protein Bax, and to a decrease in the anti-apoptotic protein Bcl-2 and in hypoxia inducible factor-1 alpha. Increased ROS generation and decreased mitochondrial membrane potential were consistent with reduced expression of antioxidative proteins identified by comparative proteomics. We therefore propose that sodium selenite induces the apoptosis of PC-3 cells mainly through the mitochondrial pathway, but also via ER stress and HIF-1 alpha mediated pathways.