Multiple Protein Analysis of Formalin-fixed and Paraffin-embedded Tissue Samples with Reverse phase Protein Arrays

被引:25
作者
Assadi, Maziar [1 ]
Lamerz, Jens [1 ]
Jarutat, Tiantom [2 ]
Farfsing, Alexandra [2 ]
Paul, Hubert [3 ]
Gierke, Berthold [4 ]
Breitinger, Ewa [4 ]
Templin, Markus F. [4 ]
Essioux, Laurent [1 ]
Arbogast, Susanne [2 ]
Venturi, Miro [3 ]
Pawlak, Michael [4 ]
Langen, Hanno [1 ]
Schindler, Thomas [1 ]
机构
[1] Hoffmann La Roche Ag, Pharma Res & Early Dev, Translat Res Sci, CH-4002 Basel, Switzerland
[2] Roche Diagnost GmbH, Pharma Res & Early Dev, Translat Res Sci, Penzberg, Germany
[3] Roche Diagnost GmbH, Pharma Res & Early Dev, Oncol, Penzberg, Germany
[4] Univ Tubingen, NMI Nat & Med Sci Inst, Dept Biochem & Prot Profiling, Reutlingen, Germany
关键词
IN-SITU HYBRIDIZATION; HER2-POSITIVE BREAST-CANCER; EXTRACTION; HER2; EXPRESSION; IMMUNOHISTOCHEMISTRY; ASSOCIATION; METHODOLOGY; PERFORMANCE; CARCINOMAS;
D O I
10.1074/mcp.M112.023051
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Reverse-phase protein arrays (RPPAs) have become an important tool for the sensitive and high-throughput detection of proteins from minute amounts of lysates from cell lines and cryopreserved tissue. The current standard method for tissue preservation in almost all hospitals worldwide is formalin fixation and paraffin embedding, and it would be highly desirable if RPPA could also be applied to formalin-fixed and paraffin embedded (FFPE) tissue. We investigated whether the analysis of FFPE tissue lysates with RPPA would result in biologically meaningful data in two independent studies. In the first study on breast cancer samples, we assessed whether a human epidermal growth factor receptor (HER) 2 score based on immunohistochemistry (IHC) could be reproduced with RPPA. The results showed very good concordance between the IHC and RPPA classifications of HER2 expression. In the second study, we profiled FFPE tumor specimens from patients with adenocarcinoma and squamous cell carcinoma in order to find new markers for differentiating these two subtypes of non-small cell lung cancer. p21-activated kinase 2 could be identified as a new differentiation marker for squamous cell carcinoma. Overall, the results demonstrate the technical feasibility and the merits of RPPA for protein expression profiling in FFPE tissue lysates.
引用
收藏
页码:2615 / 2622
页数:8
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