Overexpression of heat shock protein 27 in squamous cell carcinoma of the uterine cervix: a proteomic analysis using archival formalin-fixed, paraffin-embedded tissues
Overexpression of heat shock protein 27 in squamous cell carcinoma of the uterine cervix: a proteomic analysis using archival formalin-fixed, paraffin-embedded tissues
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DOI：
10.1016/j.humpath.2008.06.010
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发表时间：
2009-01-01
期刊：
影响因子：
3.3
通讯作者：
Tadokoro, Mamoru
中科院分区：
文献类型：
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作者：
Ono, Akiko;Kumai, Toshio;Tadokoro, Mamoru
采用福尔马林固定石蜡包埋组织的总蛋白对宫颈鳞状细胞癌进行蛋白质组学分析。使用最近开发的基于热诱导抗原修复技术的方案，从福尔马林固定的石蜡包埋组织中提取分子量为10至大于200 kd的各种蛋白质。提取的蛋白质从正常鳞状上皮（n = 53）和鳞状细胞癌（n = 21）进行荧光标记，并使用二维凝胶电泳分离。我们鉴定了728个差异表达蛋白，与正常鳞状上皮组织样本相比，144个上调，584个下调。采用液相色谱-串联质谱法分析了9种在鳞状细胞癌中表达明显上调的蛋白质。在所鉴定的候选蛋白中，用Western印迹法分析了微小染色体维持蛋白9、去整合素和金属蛋白酶结构域18以及热休克蛋白27，导致热休克蛋白27在鳞状细胞癌中显著过表达于正常粘膜（P <0.05）。此外，免疫组化显示热休克蛋白27的过度表达不仅在鳞状细胞癌，但在不同阶段的宫颈上皮内瘤变（1-3级，n = 90），包括异型增生和原位癌。热休克蛋白27在宫颈上皮内瘤变I ~ 3级和鳞癌中的表达水平显著高于正常宫颈粘膜（P <0.05）。在肿瘤性病变中，3级宫颈上皮内瘤变和鳞状细胞癌的热休克蛋白27表达水平明显高于1级宫颈上皮内瘤变（P < .05）。这些结果可能表明热休克蛋白27在宫颈上皮内瘤变-鳞状细胞癌序列中的肿瘤发生和进展中的作用。未来的实验使用福尔马林固定，石蜡包埋组织为基础的蛋白质组学分析将是一个强大的工具，为各种病理学研究。(c)2009 Elsevier Inc. All rights reserved.
Proteomic analysis of squamous cell carcinoma of the uterine cervix was performed using total protein from archival formalin-fixed, paraffin-embedded tissues. A wide range of proteins with molecular weights of 10 to greater than 200 kd was extracted from formalin-fixed, paraffin-embedded tissues using a recently developed protocol based on the heat-induced antigen retrieval technique. The extracted proteins from normal squamous epithelium (n = 53) and squamous cell carcinoma (n = 21) were fluorescently labeled and separated using 2-dimensional gel electrophoresis. We identified 728 differentially expressed proteins, with 144 up-regulated and 584 down-regulated as compared with normal squamous epithelial tissue samples. Nine proteins showing pronounced up-regulation in squamous cell carcinoma were analyzed on liquid chromatography-tandem mass spectrometry. Among the candidate proteins identified, minichromosome maintenance 9, a disintegrin and metalloproteinase domain 18, and heat shock protein 27 were analyzed in Western blotting, resulting in significant overexpression of heat shock protein 27 in squamous cell carcinoma over normal mucosa (P < .05). Furthermore, immunostaining revealed heat shock protein 27 overexpression not only in squamous cell carcinoma but in various stages of cervical intraepithelial neoplasia (grades 1-3, n = 90), including dysplasia and carcinoma in situ. The expression levels of heat shock protein 27 in cervical intraepithelial neoplasia grades I to 3 and squamous cell carcinoma were significantly higher than that in normal mucosa (P < .05). In the neoplastic lesions, heat shock protein 27 expression levels in cervical intraepithelial neoplasia grade 3 and squamous cell carcinoma were significantly higher than that in cervical intraepithelial neoplasia grade 1 (P < .05). These results may suggest a role of heat shock protein 27 in tumor development and progression in the cervical intraepithelial neoplasia-squamous cell carcinoma sequence. Future experiments using formalin-fixed, paraffin-embedded tissue-based proteomic analysis will be a powerful tool for various pathologic Studies. (c) 2009 Elsevier Inc. All rights reserved.