Protein nitration and nitrosylation by NO-donating aspirin in colon cancer cells: Relevance to its mechanism of action.
Protein nitration and nitrosylation by NO-donating aspirin in colon cancer cells: Relevance to its mechanism of action.
复制标题
DOI：
10.1016/j.yexcr.2011.03.001
复制
发表时间：
2011-06-10
影响因子：
3.7
通讯作者：
Rigas, Basil
中科院分区：
文献类型：
--
作者：
Williams, Jennie L.;Ji, Ping;Ouyang, Nengtai;Kopelovich, Levy;Rigas, Basil
关键词：
一氧化氮给药阿司匹林（NO-ASA）是一种很有前途的抗癌药物。虽然研究广泛，但其分子靶点和作用机制尚不清楚。信号蛋白的s -亚硝基化是一氧化氮的重要调控机制。在这里，我们研究了NO-ASA对NF-κB、p53和Wnt信号蛋白的s -亚硝基化是否可以部分解释其在结肠癌中的作用机制。NO- asa在HCT116和HT-29结肠细胞内释放大量的NO。通过改进的生物素开关实验，我们证明了NO-ASA s -亚硝基化结肠癌细胞中的信号蛋白p53、β-catenin和NF-κ b具有时间和浓度依赖性。NO-ASA抑制NF-κB与其同源DNA寡核苷酸的结合，而不改变NF-κB亚基p65和p50的核水平，并被二硫苏糖醇逆转，将-S-NO还原为-SH。除了s -亚硝基化外，我们还在体内和体外记录了NO-ASA对细胞蛋白酪氨酸残基的广泛硝化作用。我们的研究结果表明，NO- asa处理后细胞内NO水平的增加通过化学修饰信号级联的关键蛋白成员来调节细胞信号传导。我们推测，s -亚硝基化和酪氨酸硝化至少是NO-ASA抑制癌细胞生长的部分原因，这可能代表了no释放剂的一般作用机制。
Nitric oxide-donating aspirin (NO-ASA) is a promising agent for cancer prevention. Although studied extensively, its molecular targets and mechanism of action are still unclear. S-nitrosylation of signaling proteins is emerging as an important regulatory mechanism by NO. Here, we examined whether S-nitrosylation of the NF-κB, p53, and Wnt signaling proteins by NO-ASA might explain, in part, its mechanism of action in colon cancer. NO-ASA releases significant amounts of NO detected intracellularly in HCT116 and HT-29 colon cells. Using a modified biotin-switch assay we demonstrated that NO-ASA S-nitrosylates the signaling proteins p53, β-catenin, and NF-κB, in colon cancer cells in a time- and concentration-dependent manner. NO-ASA suppresses NF-κB binding to its cognate DNA oligonucleotide, which occurs without changes in the nuclear levels of the NF-κB subunits p65 and p50 and is reversed by dithiothreitol that reduces –S-NO to –SH. In addition to S-nitrosylation, we documented both in vitro and in vivo widespread nitration of tyrosine residues of cellular proteins in response to NO-ASA. Our results suggest that the increased intracellular NO levels following treatment with NO-ASA modulate cell signaling by chemically modifying key protein members of signaling cascades. We speculate that S-nitrosylation and tyrosine nitration are responsible, at least in part, for the inhibitory growth effect of NO-ASA on cancer cell growth and that this may represent a general mechanism of action of NO-releasing agents.
登录
查看更多内容
影响因子：
7.3
作者：
Penning, TD;Talley, JJ;Isakson, PC
通讯作者：
Isakson, PC
影响因子：
7
作者：
Park, SW;Huq, M;Wei, LN
通讯作者：
Wei, LN
影响因子：
5.1
作者：
Gu Z;Nakamura T;Lipton SA
通讯作者：
Lipton SA
影响因子：
13.6
作者：
Foster MW;Hess DT;Stamler JS
通讯作者：
Stamler JS
影响因子：
4.4
作者：
Fiorucci, S;Santucci, L;Morelli, A
通讯作者：
Morelli, A