Proteome analysis of responses to ascochlorin in a human osteosarcoma cell line by 2-D gel electrophoresis and MALDI-TOF MS
Proteome analysis of responses to ascochlorin in a human osteosarcoma cell line by 2-D gel electrophoresis and MALDI-TOF MS
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DOI：
10.1021/pr060111i
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发表时间：
2006-10-06
影响因子：
4.4
通讯作者：
Chang, Young-Chae
中科院分区：
文献类型：
--
作者：
Kang, Jeong Han;Park, Kwan-Kyu;Chang, Young-Chae
Ascochlorin是一种异戊烯基苯酚化合物，从真菌Ascochyta viciae中分离出来。抗坏血酸可降低血清胆固醇和甘油三酯水平，抑制高血压和肿瘤发展，并改善I型和II型糖尿病。在这里，为了更好地了解机制，其中子囊二氢卟酚调节生理或病理事件，并诱导癌症的药物治疗的反应，我们进行了差异分析的蛋白质组的人骨肉瘤细胞U2 OS在响应子囊二氢卟酚。此外，我们还建立了U2 OS蛋白质组的第一个二维图谱。使用双向电泳，基质辅助激光解吸/电离质谱和生物信息学的U2 OS细胞蛋白质组进行了比较，没有与子囊二氢卟酚处理。观察到表皮生长因子受体（降低4倍）、核酮糖-5-磷酸差向异构酶（降低13倍）、ATP依赖性RNA解旋酶（降低8倍）和kelch样ECH相关蛋白1（降低6倍）的表达差异最大。异质核核糖核蛋白L和微小染色体维持蛋白7的丰度分别增加了12倍和8.2倍。此外，在用子囊二氢卟酚处理的U2 OS细胞中，Erk 2增加了3倍。通过Western blotting、酶谱分析和RT-PCR分析证实了部分蛋白的表达。
Ascochlorin is a prenyl-phenol compound that was isolated from the fungus Ascochyta viciae. Ascochlorin reduces serum cholesterol and triglyceride levels, suppresses hypertension and tumor development, and ameliorates type I and II diabetes. Here, to better understand the mechanisms by which ascochlorin regulates physiological or pathological events and induces responses in the pharmacological treatment of cancer, we performed differential analysis of the proteome of the human osteosarcoma cells U2OS in response to ascochlorin. In addition, we established the first two-dimensional map of the U2OS proteome. The U2OS cell proteomes with and without treatment with ascochlorin were compared using two-dimensional electrophoresis, matrix-assisted laser desorption/ionization mass spectrometry and bioinformatics. The largest differences in expression were observed for the epidermal growth factor receptor (4-fold decrease), ribulose-5-phosphate-epimerase (13-fold decrease), ATP-dependent RNA helicase (8-fold decrease), and kelch-like ECH-associated protein 1 (6-fold decrease). The abundance of heterogeneous nuclear ribonucleoprotein L and minichromosome maintenance protein 7 increased 12- and 8.2-fold, respectively. In addition, Erk 2 was increased 3-fold in U2OS cells treated with ascochlorin. The expression of some selected proteins was confirmed by western blotting, zymography and RT-PCR analysis.