Malignant transformation in a defined genetic background: proteome changes displayed by 2D-PAGE

Molecular Cancer - Tập 9 - Trang 1-15 - 2010
Stephanie M Pütz1,2, Fotini Vogiatzi3, Thorsten Stiewe3, Albert Sickmann4
1Rudolf Virchow Center, DFG Research Center for Experimental Biomedicine, University of Würzburg, (Protein Mass Spectrometry and Functional Proteomics), Würzburg, Germany
2Institute of Medical Radiation and Cell Research (MSZ), University of Würzburg, Würzburg, Germany
3Molecular Oncology, Philipps-University Marburg, Marburg, Germany
4Leibniz-Institut für Analytische Wissenschaften - ISAS - e.V, Dortmund, Germany

Tóm tắt

Cancer arises from normal cells through the stepwise accumulation of genetic alterations. Cancer development can be studied by direct genetic manipulation within experimental models of tumorigenesis. Thereby, confusion by the genetic heterogeneity of patients can be circumvented. Moreover, identification of the critical changes that convert a pre-malignant cell into a metastatic, therapy resistant tumor cell, however, is one necessary step to develop effective and selective anti-cancer drugs. Thus, for the current study a cell culture model for malignant transformation was used: Primary human fibroblasts of the BJ strain were sequentially transduced with retroviral vectors encoding the genes for hTERT (cell line BJ-T), simian virus 40 early region (SV40 ER, cell line BJ-TE) and H-Ras V12 (cell line BJ-TER). The stepwise malignant transformation of human fibroblasts was analyzed on the protein level by differential proteome analysis. We observed 39 regulated protein spots and therein identified 67 different proteins. The strongest change of spot patterns was detected due to integration of SV40 ER. Among the proteins being significantly regulated during the malignant transformation process well known proliferating cell nuclear antigen (PCNA) as well as the chaperones mitochondrial heat shock protein 75 kDa (TRAP-1) and heat shock protein HSP90 were identified. Moreover, we find out, that TRAP-1 is already up-regulated by means of SV40 ER expression instead of H-Ras V12. Furthermore Peroxiredoxin-6 (PRDX6), Annexin A2 (p36), Plasminogen activator inhibitor 2 (PAI-2) and Keratin type II cytoskeletal 7 (CK-7) were identified to be regulated. For some protein candidates we confirmed our 2D-PAGE results by Western Blot. These findings give further hints for intriguing interactions between the p16-RB pathway, the mitochondrial chaperone network and the cytoskeleton. In summary, using a cell culture model for malignant transformation analyzed with 2D-PAGE, proteome and cellular changes can be related to defined steps of tumorigenesis.

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Molecular Cancer

Tập 9

1-15

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