Jana Čumová

Proteomic Analysis in Multiple Myeloma Research

Multiple myeloma (MM) is an incurable plasma cell (PC) malignancy characterized by the accumulation of monoclonal PCs in the bone marrow. For deeper understanding of the molecular mechanisms involved in the development of this disease, the influence of microenvironment, or the prediction of response of tumor PCs to anti-MM treatment, it is possible to use modern technologies for genomic and proteomic analyses. Due to progress in instrumentation, one of the main tools of proteomic analysis is mass spectrometry in combination with chosen separation techniques. This review will provide a short survey of the most commonly used proteomic techniques and show examples of their applications in MM proteome studies.

Bone marrow plasma cell separation - validation of separation algorithm

Monoclonal gammopathy is characterized by the presence of monoclonal immunoglobulin which is produced by a specifi c clone of terminally differentiated long-living plasma cells (PC) in the bone marrow. More detailed understanding of this clone at the genomic and proteomic level as well as its comparison with the population of normal PC is possible only after the clone is separated from other BM cells.

B022 The Effect of Bortezomib on Myeloma Cell Line

Introduction: Multiple myeloma (MM) is still an incurable disease characterized by the clonal expansion of malignant plasma cells. New anticancer drugs further improve prognosis of myeloma patients. The aim of this study was to evaluate changes in protein expression of myeloma cell line ARH 77 after bortezomib treatment. Materials and methods: Myeloma cell line ARH 77 was treated with bortezomib (10 – 40nM) for various periods of time (24 and 48 hours). The proteins contained in total myeloma cell lysate were separated by 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and the differentially expressed proteins between the untreated and treated cell lines were excised and identified by mass spectrometry. Results: There were analyzed 94 proteins differentially expressed between treated and control cells; total of 34 protein spots were upregulated: proteins involved in regulation of apoptosis, chaperons/stress related proteins, proteolysis of ubiquitin/protein degradation and cytoskeleton proteins. Sixty protein spots were downregulated: proteins involved in synthesis, regulation of apoptosis, chaperons/stress related proteins, regulation of cell cycle proteins, proteins connected to glycolysis and proteolysis of ubiquitin/protein degradation and antioxidant/redox proteins. Conclusion: We identified 94 proteins altered in myeloma cells after various exposure of to bortezomib. This proteomic approach can contribute to elucidation of mechanisms of new anticancer drugs action.