Validation of a robust proteomic analysis carried out on formalin-fixed paraffin-embedded tissues of the pancreas obtained from mouse and human.
Validation of a robust proteomic analysis carried out on formalin-fixed paraffin-embedded tissues of the pancreas obtained from mouse and human.
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DOI：
10.1002/pmic.201100663
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发表时间：
2012-11
期刊：
影响因子：
3.4
通讯作者：
Mobley, James A.
中科院分区：
文献类型：
--
作者：
Kojima, Kyoko;Bowersock, Gregory J.;Kojima, Chinatsu;Klug, Christopher A.;Grizzle, William E.;Mobley, James A.
最近出现了一些报告，重点是从福尔马林固定石蜡包埋（FFPE）组织中提取蛋白质用于MS分析;然而，与快速冷冻对照相比，重现性和耐用性通常被忽视。本研究的目的是确定和验证一个实用的和高度稳健的FFPE组织的蛋白质组学分析方法。使用1D-nanoLC-MS（MS）2分析从小鼠（n = 8）获得的FFPE和匹配的冷冻胰腺组织，随后使用市售试剂盒进行处理。发现FFPE组织的所选方法与冷冻方法具有高度可比性。此外，两组之间鉴定的独特肽的总数高度相似，FFPE鉴定了958个，冷冻鉴定了1070个，蛋白质鉴定对应约80%。然后将该方法应用于存档的人FFPE胰腺癌标本（n = 11），与未涉及的组织（n = 8）相比，其中47种潜在的胰腺导管腺癌标志物被鉴定为显著增加，其中28种是先前报道的。此外，这些蛋白质与胰腺癌（包括雌激素受体α）具有强烈重叠的通路相关性。总之，这些数据支持验证FFPE组织的蛋白质组学分析方法，该方法简单且高度稳健，也可有效应用于疾病的转化研究。
A number of reports have recently emerged with focus on extraction of proteins from formalin-fixed paraffin-embedded (FFPE) tissues for MS analysis; however, reproducibility and robustness as compared to flash frozen controls is generally overlooked. The goal of this study was to identify and validate a practical and highly robust approach for the proteomics analysis of FFPE tissues. FFPE and matched frozen pancreatic tissues obtained from mice (n = 8) were analyzed using 1D-nanoLC-MS(MS)2 following work up with commercially available kits. The chosen approach for FFPE tissues was found to be highly comparable to that of frozen. In addition, the total number of unique peptides identified between the two groups was highly similar, with 958 identified for FFPE and 1070 identified for frozen, with protein identifications that corresponded by approximately 80%. This approach was then applied to archived human FFPE pancreatic cancer specimens (n = 11) as compared to uninvolved tissues (n = 8), where 47 potential pancreatic ductal adenocarcinoma markers were identified as significantly increased, of which 28 were previously reported. Further, these proteins share strongly overlapping pathway associations to pancreatic cancer that include estrogen receptor α. Together, these data support the validation of an approach for the proteomic analysis of FFPE tissues that is straightforward and highly robust, which can also be effectively applied toward translational studies of disease.
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作者：
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