Identification of novel nasopharyngeal carcinoma biomarkers by laser capture microdissection and proteomic analysis
Identification of novel nasopharyngeal carcinoma biomarkers by laser capture microdissection and proteomic analysis
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DOI：
10.1158/1078-0432.ccr-07-1215
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发表时间：
2008-01-15
影响因子：
11.5
通讯作者：
Xiao, Zhi-Qiang
中科院分区：
文献类型：
--
作者：
Cheng, Ai-Lan;Huang, Wei-Guo;Xiao, Zhi-Qiang
目的：采用激光捕获显微切割技术和蛋白质组学方法鉴定鼻咽癌（NPC）新的生物标志物。实验设计：采用双向凝胶电泳技术分离NPC和正常鼻咽上皮组织（NNET）的蛋白质，并通过质谱鉴定差异蛋白。用Western blotting法检测上述两种组织和四种鼻咽癌细胞系中三种差异蛋白（stathmin、14-3-3 sigma和annexin I）的表达。应用免疫组化法检测98例原发性NPC、30例NNET和20例颈淋巴结转移瘤组织中3种差异蛋白的表达，并分析其表达水平与临床病理特征和临床预后的关系。证实了两种组织中stathmin、14-3-3 sigma和annexin I的表达水平与NPC细胞系的分化程度和/或转移潜能相关。与NNET相比，在NPC中观察到14-3-3 sigma和膜联蛋白I的显著stathmin上调和下调，并且与原发性NPC相比，在淋巴结转移中也观察到14-3-3 sigma和膜联蛋白I的显著下调。此外，stathmin上调和下调14-3-3 sigma和膜联蛋白I与组织学分化差、晚期临床分期和复发显著相关，而下调14-3-3 sigma和膜联蛋白I也与淋巴结和远处转移显著相关。此外，生存曲线显示，stathmin上调和14-3-3 sigma和膜联蛋白I下调的患者预后不良。多因素分析显示stathmin、14-3-3 sigma和annexin I的表达状态是独立的预后指标。结论：stathmin、14-3-3 sigma和annexin I是NPC分化和预后的潜在生物标志物，其表达异常可能在NPC的发病机制中起重要作用。
Purpose: To identify novel nasopharyngeal carcinoma (NPC) biomarkers by laser capture microdissection and a proteomic approach.Experimental Design: Proteins from pooled microdissected NPC and normal nasopharyngeal epithelial tissues (NNET) were separated by two-dimensional gel electrophoresis, and differential proteins were identified by mass spectrometry. Expression of three differential proteins (stathmin, 14-3-3 sigma, and annexin I) in the above two tissues as well as four NPC cell lines was determined by Western blotting. Immunohistochemistry was also done to detect the expression of three differential proteins in 98 cases of primary NPC, 30 cases of NNET, and 20 cases of cervical lymph node metastases, and the correlation of their expression levels with clinicopathologic features and clinical outcomes were evaluated.Results: Thirty-six differential proteins between the NPC and NNET were identified. The expression levels of stathmin,14-3-3 sigma, and annexin I in the two types of tissues were confirmed and related to differentiation degree and/or metastatic potential of the NPC cell lines. Significant stathmin up-regulation and down-regulation of 14-3-3 sigma and annexin I were observed in NPC versus NNET, and significant down-regulation of 14-3-3 sigma and annexin I was also observed in lymph node metastasis versus primary NPC. In addition, stathmin up-regulation and down-regulation of 14-3-3 sigma and annexin I were significantly correlated with poor histologic differentiation, advanced clinical stage, and recurrence, whereas down-regulation of 14-3-3 sigma and annexin I was also significantly correlated with lymph node and distant metastasis. Furthermore, survival curves showed that patients with stathmin up-regulation and down-regulation of 14-3-3 sigma and annexin I had a poor prognosis. Multivariate analysis revealed that the expression status of stathmin,14-3-3 sigma, and annexin I was an independent prognostic indicator.Conclusion: The data suggest that stathmin,14-3-3 sigma, and annexin I are potential biomarkers for the differentiation and prognosis of NPC, and their dysregulation might play an important role in the pathogenesis of NPC.