Expression profiling of breast cancer cells by differential peptide display |
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Authors: | Tammen Harald Kreipe Hans Hess Rüdiger Kellmann Markus Lehmann Ulrich Pich Andreas Lamping Norbert Schulz-Knappe Peter Zucht Hans-Dieter Lilischkis Richard |
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Institution: | (1) BioVisioN AG, Medizinische Hochschule Hannover, Hannover;(2) Institute of Pathology, Medizinische Hochschule Hannover, Hannover;(3) Institute of Biochemistry, Klinikum RWTH, Germany |
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Abstract: | Expression profiling of RNAs or proteins has become a promising means to investigate the heterogeneity of histopathologically defined classes of cancer. Peptides, representing degradation as well as processing products of proteins offer an even closer insight into cell physiology. Peptides are related to the turnover of cellular proteins and are capable to reflect disease-related changes in homoeostasis of the human body. Furthermore, peptides derived from tumor cells are potentially useful markers in the early detection of cancer.In this study, we introduced a method called differential peptide display (DPD) for separating, detecting, and identifying native peptides derived from whole cell extracts. This method is a highly standardized procedure, combining the power of reversed-phase chromatography with mass spectrometry. This technology is suitable to analyze cell lines, various tissue types and human body fluids. Peptide-based profiling of normal human mammary epithelial cells (HMEC) and the breast cancer cell line MCF-7 revealed complex peptide patterns comprising of up to 2300 peptides. Most of these peptides were common to both cell lines whereas about 8% differed in their abundance. Several of the differentially expressed peptides were identified as fragments of known proteins such as intermediate filament proteins, thymosins or Cathepsin D. Comparing cell lines with native tumors, overlapping peptide patterns were found between HMEC and a phylloides tumor (CP) on the one hand and MCF-7 cells and tissue from a invasive ductal carcinoma (DC) on the other hand. |
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Keywords: | breast cancer differential peptide display expression profiling MALDI mass spectrometry proteomics reversed-phase HPLC |
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