Chao Jui-I, Kuo Pao-Chen, Hsu Tzu-Sheng
Molecular Toxicology Laboratory, Institute of Pharmacology and Toxicology, College of Life Sciences, Tzu Chi University, 701 Section 3 Chung-Yang Road, Hualien 970, Taiwan.
J Biol Chem. 2004 May 7;279(19):20267-76. doi: 10.1074/jbc.M312381200. Epub 2004 Feb 26.
Survivin is expressed in most tumor cells and has been associated with both anti-apoptosis and mitotic progression. However, the mechanism of regulation of the survivin expression remains unclear. In this study we investigated the expression and regulation of survivin in the nitric oxide (NO)-exposed human lung carcinoma cells. The lung carcinoma cell lines CL3, H1299, and A549 but not normal lung fibroblast expressed high levels of survivin proteins. NO donors S-nitroso-N-acetyl-penicillamine (SNAP) and sodium nitroprusside (SNP) decreased the survivin expression. SNAP (0.4 mm, 24h)and SNP (1 mm, 24 h) significantly induced cytotoxicity and apoptosis in lung carcinoma cells. Furthermore, SNAP inhibited the cell growth and increased the fractions of G(2)/M phase. The levels of cyclin B1 and phospho-cdc2-(Thr-161) proteins were inhibited in the NO-exposed cells. The cdc25 phosphatase inhibitors (Cpd 5 and NSC 663284) and the cdc2 kinase inhibitors (alsterpaullone and purvalanol A) enhanced SNP-induced cytotoxicity and the decrease in survivin expression. However, overexpression of survivin by a pOTB7-survivin vector reduced SNP-induced cell growth inhibition and cytotoxicity. In addition, SNP activated the phosphorylation of p38 mitogen-activated protein (MAP) kinase. The specific p38 MAP kinase inhibitor, SB202190, significantly decreased the cytotoxicity and increased the survivin levels in NO donor-treated and inducible NOS-transfected cells. Conversely, anticancer agents including quercetin, arsenite, and cisplatin but not genistein increased the levels of survivin protein. Our results indicated for the first time that NO inhibited the expression of survivin, which was down-regulated by the p38 MAP kinase pathway.
生存素在大多数肿瘤细胞中表达,并与抗凋亡和有丝分裂进程相关。然而,生存素表达的调控机制仍不清楚。在本研究中,我们调查了一氧化氮(NO)处理的人肺癌细胞中生存素的表达和调控情况。肺癌细胞系CL3、H1299和A549而非正常肺成纤维细胞表达高水平的生存素蛋白。NO供体S-亚硝基-N-乙酰青霉胺(SNAP)和硝普钠(SNP)降低了生存素的表达。SNAP(0.4 mM,24小时)和SNP(1 mM,24小时)显著诱导肺癌细胞的细胞毒性和凋亡。此外,SNAP抑制细胞生长并增加G(2)/M期细胞比例。在NO处理的细胞中,细胞周期蛋白B1和磷酸化cdc2-(苏氨酸161)蛋白水平受到抑制。cdc25磷酸酶抑制剂(化合物5和NSC 663284)和cdc2激酶抑制剂(阿尔斯特帕隆和嘌呤瓦诺A)增强了SNP诱导的细胞毒性以及生存素表达的降低。然而,通过pOTB7-生存素载体过表达生存素可降低SNP诱导的细胞生长抑制和细胞毒性。此外,SNP激活了p38丝裂原活化蛋白(MAP)激酶的磷酸化。特异性p38 MAP激酶抑制剂SB202190显著降低了NO供体处理细胞和诱导型一氧化氮合酶转染细胞中的细胞毒性,并增加了生存素水平。相反,包括槲皮素、亚砷酸盐和顺铂但不包括染料木黄酮在内的抗癌药物增加了生存素蛋白水平。我们的结果首次表明,NO抑制生存素的表达,而生存素表达是由p38 MAP激酶途径下调的。