Towards understanding the mechanisms of metastasis in low grade breast cancer: Discovery of new targets with proteomics-driven transcriptomic approach
| Autoři | |
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| Rok publikování | 2011 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | Although breast cancer is the most common women cancer in the world, traditional prognostic factors are not sufficient for precise risk-group discrimination. This is well illustrated by the group of low grade breast tumors in which small percentage of tumors form lymph node metastases early with very bad response to the therapy. Our effort has been focused on biomarker discovery in the set of 96 clinicopathologically well characterized set of breast cancer primary tumors grouped and randomized according to grade, lymph node and hormonal receptor status. A high resoltution proteomics approaches involving iTRAQ labelling, two-dimensional chromatography and high resolution mass spectrometry (iTRAQ-2DLC-MS/MS) together with parallel 2-D gel electrophoresis were involved in complex protein analysis of tissue lysates. Subsequently, we designed a 384-well TaqMan qRT-PCR array covering the most promising targets identified by proteomics approach (based on their expression profile and biological role) which also involved another metastasis-related genes not detected at the protein level to obtain gene expression profiles at transcript level. Immunohistochemical staining of tissue arrays is currently in progress to confirm the protein levels of most important targets at tissue level. Our results indicate several types of differences between the studied groups of tumors, namely: (i) iTRAQ-2DLC-MS/MS data were internally validated via ErbB2 protein expression as confirmed by immunohistochemistry and by strong induction of AGR family proteins in estrogene receptor positive samples. (ii) Biological replicates in the study exhibited strong positive correlation and clustering of proteomic profiles according to clinicopathological parameters. (iii) Expression of selected cytoskeletal proteins, proteases and NF-kappaB regulated proteins correlated with lymph node status of the tumors. Our findings provide a solid data for targeted functional studies and validation geared towards more in-depth description of metastatic mechanisms in low-grade breast cancer tumors. |
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