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Project Q

Project Q

 

 

Name:

Prof. Dr. Brigitte Schlegelberger

 

Institution:

Institute of Cell- and Molecular Pathology

 

Telephone:

0049-511-532-4522

 

Email:

Schlegelberger.Brigittemh-hannover.de

 

Projects:

Molecular characterization of transgenic breast cancer mouse models In cooperation with the group of Wolfgang Deppert (Heinrich-Pette-Institut Hamburg) it is planned to characterize the WAP-T and WAP-T-NP lines generated by this group (Schulze-Garg C, Löhler J, Gocht A, Deppert W, Oncogene 19, 1028-1037, 2000). In these mice, the SV40 early genomic region is driven by the WAP-promotor that is developmentally regulated by lactotrophic hormones. Thus, the oncogenic large-T-antigen that inactivates the tumor suppressor proteins Rb and p53 is specifically induced in epithelial cells of the terminal duct lobular units in lactating mice. They develop multifocal DCIS and consequently invasive breast carcinomas within strain specific periods of latency. DCIS lesions in transgenic mice exhibit architectural and cytological features which closely resemble those commonly present in humans. In the first phase of this project, the serum of the WAP-T and WAP-T-NP mice and the serum of wild-type mice will be analysed at defined time points by means of the chip-based protein-profiling technology surface-enhanced laser desorption/ ionization (SELDI-MS). After protein extraction with established methods and purification with HPLC, SELDI-specific protein chips with different surfaces will be used to show specific protein signals. The different surfaces of the chips allow the analysis of cationic, anionic, hydrophobic, and non-hydrophobic protein structures. It is also possible to catch specific proteins after labeling the surface of the chips with DNA or antibodies. Different matrices and the changes in laser intensity or in the detection sensitivity make it possible to look for peptides and proteins in the range between 1 and 20 kd. The pattern will be compared with internationally available bioinformatic tools such as ProFound (http://www.prowl.rockefeller.edu/). It will thus be possible to identify differentially expressed proteins and post-translational protein modifications. In addition, sequencing of the proteins is possible with a MALDI-TOF device. The protein profiles identified in WAP-T and WAP-T-NP mice will be compared with those of other transgenic mouse model like the MTV-induced breast carcinomas and with protein profiles of patients with breast cancer and age-matched controls. Of special interest are proteins expressed during early phases of breast cancer development. Differentially expressed proteins will be validated by alternative methods, e.g. immunohistochemistry or Western blotting In the second phase of the project, cytogenetic studies will be performed to understand the role of chromosome aberrations during tumor development. Fluorescence- in situ-hybridisation with self-generated DNA-probes binding to chromosome regions frequently involved in breast cancer will make it possible to detect deletions and amplifications. If we will succeed to prepare metaphases, spectral karyotyping (SKY) will be applied. As soon as matrix-CGH-chips for the mouse genome are available, they will be used to determine chromosomal losses and gains, especially amplifications most precisely. This project opens the chance to investigate early changes during breast cancer development, and to identify new prognostic markers.

 

Group members:

Prof. Dr. med. Brigitte Schlegelberger

Dr. rer. nat. Nils von Neuhoff

Dr. rer. nat. Doris Steinmann

 

Methods:

Chromosome analysis

Fluorescence in situ hybridization

Spectral karyotyping

Generation and labelling of BACs and cosmids

DNA-sequencing

Microsatellite analysis

Cell culture techniques

 

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