采用液相色谱-串联质谱技术对福尔马林固定石蜡包埋的胰腺组织进行蛋白质组学分析。
Proteomic analysis of formalin-fixed paraffin-embedded pancreatic tissue using liquid chromatography tandem mass spectrometry.
机构信息
Department of Pathology, Harvard Medical School, Boston, MA, USA.
出版信息
Pancreas. 2012 Mar;41(2):175-85. doi: 10.1097/MPA.0b013e318227a6b7.
OBJECTIVES
FFPE tissue is a standard method of specimen preservation for hospital pathology departments. Formalin-fixed paraffin-embedded tissue banks are a resource of histologically characterized specimens for retrospective biomarker investigation. We aim to establish liquid chromatography coupled with tandem mass spectrometry analysis of FFPE pancreatic tissue as a suitable strategy for the study of the pancreas proteome.
METHODS
We investigated the proteomic profile of FFPE pancreatic tissue specimens, using liquid chromatography coupled with tandem mass spectrometry, from 9 archived specimens that were histologically classified as normal (n = 3), chronic pancreatitis (n = 3), and pancreatic cancer (n = 3).
RESULTS
We identified 525 nonredundant proteins from 9 specimens. Implementing our filtering criteria, 78, 15, and 21 proteins were identified exclusively in normal, chronic pancreatitis, and pancreatic cancer specimens, respectively. Several proteins were identified exclusively in specimens with no pancreatic disease: spink 1, retinol dehydrogenase, and common pancreatic enzymes. Similarly, proteins were identified exclusively in chronic pancreatitis specimens: collagen α1 (XIV), filamin A, collagen α3 (VI), and SNC73. Proteins identified exclusively in pancreatic cancer included annexin 4A and fibronectin.
CONCLUSIONS
We report that differentially expressed proteins can be identified among FFPE tissue specimens originating from individuals with different pancreatic histologic findings. The mass spectrometry-based method used herein has the potential to enhance biomarker discovery and chronic pancreatitis research.
目的
FFPE 组织是医院病理科标本保存的标准方法。福尔马林固定石蜡包埋组织库是用于回顾性生物标志物研究的具有组织学特征标本的资源。我们旨在建立 FFPE 胰腺组织的液相色谱串联质谱分析,作为研究胰腺蛋白质组的合适策略。
方法
我们使用液相色谱串联质谱法研究了 9 份存档的 FFPE 胰腺组织标本的蛋白质组学图谱,这些标本在组织学上被分类为正常(n = 3)、慢性胰腺炎(n = 3)和胰腺癌(n = 3)。
结果
我们从 9 个标本中鉴定出 525 个非冗余蛋白。实施我们的过滤标准后,分别在正常、慢性胰腺炎和胰腺癌标本中鉴定出 78、15 和 21 种蛋白质是特异性的。一些蛋白质仅在没有胰腺疾病的标本中被鉴定出来:spink1、视黄醇脱氢酶和常见的胰腺酶。同样,在慢性胰腺炎标本中鉴定出了特异性的蛋白质:胶原 α1(XIV)、细丝蛋白 A、胶原 α3(VI)和 SNC73。在胰腺癌标本中鉴定出的蛋白质包括膜联蛋白 4A 和纤维连接蛋白。
结论
我们报告说,可以在源自具有不同胰腺组织学发现的个体的 FFPE 组织标本中鉴定出差异表达的蛋白质。本文中使用的基于质谱的方法有可能增强生物标志物的发现和慢性胰腺炎的研究。