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Comparative proteomic analysis of normal and tumor stromal cells by tissue on chip based mass spectrometry (toc-MS)

Niko Escher1, Günther Ernst1, Christian Melle1, Alexander Berndt2, Joachim H Clement3, Kerstin Junker4, Karlheinz Friedrich5, Orlando Guntinas-Lichius6 and Ferdinand von Eggeling1*

Author affiliations

1 Core Unit Chip Application, Institute of Human Genetics, University Hospital Jena, 07740 Jena, Germany

2 Institute of Pathology, University Hospital Jena, 07740 Jena, Germany

3 Department of Hematology and Oncology, Clinic for Internal Medicine II, University Hospital Jena, 07740 Jena, Germany

4 Department of Urology, University Hospital Jena, Lessingstrasse 1, 07740 Jena, Germany

5 Institute of Biochemistry II, University Hospital Jena, Nonnenplan 2, 07740 Jena, Germany

6 Department of Otorhinolaryngology, University Hospital Jena, 07740 Jena, Germany

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Citation and License

Diagnostic Pathology 2010, 5:10  doi:10.1186/1746-1596-5-10

Published: 28 January 2010

Abstract

In carcinoma tissues, genetic and metabolic changes not only occur at the tumor cell level, but also in the surrounding stroma. This carcinoma-reactive stromal tissue is heterogeneous and consists e.g. of non-epithelial cells such as fibroblasts or fibrocytes, inflammatory cells and vasculature-related cells, which promote carcinoma growth and progression of carcinomas. Nevertheless, there is just little knowledge about the proteomic changes from normal connective tissue to tumor stroma. In the present study, we acquired and analysed specific protein patterns of small stromal sections surrounding head and neck cell complexes in comparison to normal subepithelial connective tissue. To gain defined stromal areas we used laser-based tissue microdissection. Because these stromal areas are limited in size we established the highly sensitive 'tissue on chip based mass spectrometry' (toc-MS). Therefore, the dissected areas were directly transferred to chromatographic arrays and the proteomic profiles were subsequently analysed with mass spectrometry. At least 100 cells were needed for an adequate spectrum. The locating of differentially expressed proteins enables a precise separation of normal and tumor stroma. The newly described toc-MS technology allows an initial insight into proteomic differences between small numbers of exactly defined cells from normal and tumor stroma.