Dr. Henning Feist
Institute of Pathology, Medical School Hannover
Invasive breast carcinoma. Intraductal breast carcinoma. Intraductal breast lesions.
In the development of malignant tumors histologically identifiable pre-invasive lesions can be observed. In operative breast specimens intraductal, i. e. non-invasive pathological changes are detectable, but for the histopathologist often it is difficult to discriminate hyperproliferative from neoplastic intraductal lesions as possible precursors of invasive breast carcinoma, especially ductal invasive carcinoma, as the most common malignant tumor of women.
Histologically ductal hyperplasia (DH), atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS) are defined. In order to investigate these lesions concerning genetic aberrations which may help to distinguish between these pathologic changes with different prognostic impact we combine laser-assisted microdissection of histologic slides with PCR-based molecularbiological methods. In this project laser-assisted microdissection is performed in order to guaranteee isolation of often small lesions without contamination, because contamination would render false results in the following performed assays.
Molecularbiological tools are PCR-based deletion-analysis of chromosomal areas (LOH) and methylation-analysis of several tumor suppressor genes via bisulfit-conversation, PCR amplification and sequencing of the microdissected material. Moreover, quantitative assessment of methylation and gene expression is possible by using real time PCR (TaqMan).
Comparing intraductal with invasive carcinoma we hope to get insights into clonal development of breast cancer isolating and analyzing small identifiable lesions we hope to improve understanding of the malignant potential of different intraductal pathologic entities.
Prof. Dr. H. H. Kreipe
Dr. H. Feist
Dr. U. Lehmann
Dr. S. Glöckner
Laser-assisted microdissection; Deletion analysis (Loss of heterozygosity); Bisulfit conversation of DNA for methylation analysis; DNA-sequencing; Real-Time PCR or quantitation of expression and methylation
Feist H, Lilischkis R, Hallas C, Aanesen JJ, Gassel A, Gallert KC, Kreipe H. Detection of clonality by laser-microdissection and PCR. Verh Dtsch Ges Path. 1997; 81:339-342 Glöckner S, Lehmann U, Wilke N, Kleeberger W, Länger F, Kreipe H. Amplification of growth regulatory genes in intraductal breast cancer is associated with higher nuclear grade but not with the progression to invasiveness. L Invest.2001;81:565-71
Lehmann U, Glöckner S, Kleeberger W, von Wasielewski R, Feist H, Kreipe H. Detection of gene amplification in archival breast cancer specimens by laser-assisted microdissection and quantitative real-time polymerase chain reaction. Am J Pathol. 2000;156:1855-64 Lehmann U, Hasemeier B, Lilischkis R, Kreipe H. Quantitative analysis of promoter hypermethylation in laser-microdissected archival specimens. Lab Invest. 2001;81:635-38