Proteomic identification of tumor biomarkers associated with primary gallbladder cancer
Proteomic identification of tumor biomarkers associated with primary gallbladder cancer
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DOI：
10.3748/wjg.v20.i18.5511
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发表时间：
2014-05-14
影响因子：
4.3
通讯作者：
Jin, Kai-Zhou
中科院分区：
文献类型：
--
作者：
Huang, Hai-Lin;Yao, Hou-Shan;Jin, Kai-Zhou
目的：目的：寻找原发性胆囊癌（PGC）的潜在生物标志物。方法：分别收集10例患者的新鲜PGC、胆囊炎和正常胆囊组织标本，进行比较蛋白质组学分析。采用双向凝胶电泳（2-DE）技术对胆囊癌、胆囊炎及正常胆囊组织的蛋白质组进行比较。然后使用MALDI-TOF质谱仪（MS）和数据库搜索鉴定差异表达的蛋白质。结果：通过2-艾德分析，在PGC、胆囊炎和正常胆囊组织中发现7个差异表达蛋白点。使用MALDI-TOF MS鉴定了7个差异表达蛋白中的6个，其中3个在PGC组织中过表达，3个在PGC组织中低表达。蛋白质印迹分析显示，与正常肿瘤相邻组织相比，PGC组织中膜联蛋白A4（ANXA 4）的蛋白水平显著升高，热休克蛋白90-β（Hsp 90 β）和动力蛋白胞质1重链1（Dync 1h 1）降低。然而，肌动蛋白，主动脉平滑肌和γ-肌动蛋白水平不变。此外，5种蛋白质的mRNA水平与蛋白质水平变化相似（P < 0.01）。免疫组化结果显示，与癌旁正常组织相比，PGC组织中ANXA 4的表达上调，Hsp 90 β和Dync 1h 1的表达下调。这些蛋白在PGC癌变过程中的作用有待进一步研究。(C)2014百世登出版集团有限公司有限公司All rights reserved.
AIM: To identify potential biomarkers of primary gallbladder cancer (PGC).METHODS: Fresh PGC, cholecystitis and normal gallbladder tissue specimens collected from 10 patients, respectively, were subjected to comparative proteomic analysis. The proteomic patterns of PGC were compared with those of cholecystitis and normal gallbladder tissues using two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were then identified using a MALDI-TOF mass spectrometer (MS) and database searches. To further validate these proteins, 20 samples of PGC tissues and normal tumor-adjacent tissues were collected for Western blot, quantitative real-time PCR, and immunohistochemical staining assay.RESULTS: Seven differentially expressed protein spots were detected by 2-ED analysis by comparing the average maps of PGC, cholecystitis and normal gallbladder tissues. Six of the seven differentially expressed proteins were identified using MALDI-TOF MS, with three overexpressed and three underexpressed in PGC tissue. Protein levels of annexin A4 (ANXA4) were significantly elevated, and heat shock protein 90-beta (Hsp90 beta) and dynein cytoplasmic 1 heavy chain 1 (Dync1h1) were decreased in PGC tissues relative to the normal tumor-adjacent tissues as shown by Western blot analysis. However, levels of actin, aortic smooth muscle and gamma-actin were unchanged. In addition, the mRNA levels of all 5 proteins showed similar changes to those of the protein levels (P < 0.01). Further validation by immunohistochemical analysis showed the upregulated expression of ANXA4 and decreased expression of Hsp90 beta and Dync1h1 in the cytoplasm of PGC tissues relative to the normal tumor-adjacent tissues.CONCLUSION: Three proteins are identified as potential biomarkers of PGC using proteomic analysis. The functions of these proteins in the carcinogenesis of PGC remain to be studied. (C) 2014 Baishideng Publishing Group Co., Limited. All rights reserved.