Isoform-specific expression and characterization of 14-3-3 proteins in human glioma tissues discovered by stable isotope labeling with amino acids in cell culture-based proteomic analysis
Isoform-specific expression and characterization of 14-3-3 proteins in human glioma tissues discovered by stable isotope labeling with amino acids in cell culture-based proteomic analysis
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DOI：
10.1002/prca.200800198
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发表时间：
2009-06-01
影响因子：
2
通讯作者：
Wei, Yuquan
中科院分区：
文献类型：
--
作者：
Liang, Shufang;Shen, Guobo;Wei, Yuquan
人14-3-3蛋白在不同类型的正常或肿瘤细胞和组织中具有异构体特异性表达和功能。然而，14-3-3蛋白在人脑胶质瘤中的表达谱及其亚型特异性的生物学功能尚不清楚。在我们的研究中，14-3-3异构体在人脑胶质瘤组织中的表达水平和特性进行了研究，通过一个敏感的，准确的稳定同位素标记的氨基酸在细胞培养为基础的定量蛋白质组学策略。结果表明，除14-3-3 σ不表达外，其余6种亚型在胶质瘤组织和癌旁脑组织中均有不同程度的表达。胶质瘤组织中14-3-3 β和eta表达上调，而14-3-3 zeta表达下调。另外三种亚型14-3-3 β、θ和γ在人脑胶质瘤组织和外周血淋巴细胞中的表达水平相似。Western blot和免疫组化分析均与定量蛋白质组学数据一致。甲基化特异性PCR分析进一步发现14-3-3 sigma基因在胶质瘤中的表达缺失是由于其编码区的DNA高度甲基化所致。这些结果提示14-3-3 β、eta、zeta和sigma四种亚型可能在人脑胶质瘤的发生发展中起重要作用，有可能作为人脑胶质瘤诊断和治疗的潜在生物标志物。
Human 14-3-3 proteins have isoform-specific expression and functions in different kinds of normal or tumor cells and tissues. However, the expression profiling of 14-3-3 proteins and isoform-specific biological functions are unclear in human glioma so far. In our study, the expression levels and characterization of 14-3-3 isoforms in human glioma tissues were investigated by a sensitive, accurate stable isotope labeling with amino acids in cell culture-based quantitative proteomic strategy. As a result, except unexpressed 14-3-3 sigma, the other six isoforms, with different expression levels, were existed in glioma tissues and para-cancerous brain tissues (PBTs). 14-3-3 beta and eta were upregulated, whereas 14-3-3 zeta was downregulated in glioma tissues compared with that in PBTs. And the other three isoforms 14-3-3 epsilon, theta, and gamma had similar expression levels in human glioma tissues and PBTs. Western blot and immunohistochemistry analysis were both consistent with the quantitative proteomic data. The loss of expression of 14-3-3 sigma was further discovered due to DNA high methylation in its coding region in glioma by methylation-specific PCR analysis. These results indicated that the four isoforms, including 14-3-3 beta, eta, zeta, and sigma, may play important roles in tumorigenesis of human glioma, which is probably used as potential biomarkers for diagnosis and targets for treatment of human gliomas in future.