A novel panel of proteins correlating with lymph node metastasis of low-grade breast cancer: Towards functional characterization and clinical validation
| Autoři | |
|---|---|
| Rok publikování | 2013 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | In our previous experiments, we have identified a panel of proteins correlating with lymph node status (N1-2 vs. N0) in small (T1) low grade (G1) breast tumors. Shortly, we have analyzed a set of 96 well characterized breast tumors designed according to grade, lymph node status and status of hormonal receptors. The discovery phase involved proteomics based on iTRAQ-2DLC-MS/MS method, transcriptomics and immunohistochemistry. A panel of identified proteins involved activators of NF-kappaB pathway, proteases and adhesive and cytoskeletal proteins. Analysis of biomarker selectivity based on agreement between proteomics, transcriptomics and immunohistochemical data divided the potential biomarkers into two selectivity groups. For most potential biomarkers, statistically significant correlation with patient survival in a completely independent set of 214 breast tumors was obtained. Cell surface proteins are important therapeutics targets in general and could serve as potential targets of anti-metastatic therapy here. Our effort thus aimed to identify novel cell surface proteins with transmembrane domains in diferrentially migrating variants of MDA-MB-231 breast cancer cell line. Using cell surface biotinylation and SILAC quantitative analysis, we identified 38/91 transmembrane proteins up-/down regulated in more migrating variant of the cell line. Five selected proteins complemented the panel of metastasis associated proteins as identified above. Our ongoing effort is oriented towards further characterizing of the panel in term of their role in pro-metastatic mechanisms. We will show the current status of our research in characterizing proteins’ role in cell migration and invasivity in vitro. Moreover, analysis of protein-protein interactions can further contribute to understanding the molecular role of particular proteins and we will show our progress in this field. Additional aim of our research project is a clinical validation of the panel using larger independent sample set. For this puprose, we have established methods for targeted proteomics quantification of all the panel protein members from FFPE archive tissue material. Moreover, next generation proteomics based on SWATH method enables generation of digital fingerprints of the proteome. This approach is believed to establish a novel concept of “digitized biobanking”. Our first data indicating applicability of this method in breast cancer research and its molecular classification will be analyzed and discussed. |