PMID- 16426913 OWN - NLM STAT- MEDLINE DCOM- 20060224 LR - 20061115 IS - 0046-8177 (Print) IS - 0046-8177 (Linking) VI - 37 IP - 2 DP - 2006 Feb TI - Improved clonality analysis of multifocal bladder tumors by combination of histopathologic organ mapping, loss of heterozygosity, fluorescence in situ hybridization, and p53 analyses. PG - 143-51 AB - The clonality status of multifocal bladder tumors is still controversially discussed with experimental evidence for both monoclonality and field cancerization. Methodologically, loss of heterozygosity (LOH) and genomic sequencing analyses are widely used in clonality analysis of malignant tumors. In the present study, we used LOH analysis and genomic sequencing in combination with fluorescence in situ hybridization (FISH) and extensive histopathologic whole-organ mapping to determine the clonal relationship of multifocal bladder cancer disease. Tissue sections (1 cm(2)) covering the entire urothelial lining were systematically dissected from 2 cystectomy specimens (cystectomy 1, no urothelial lesions, bladder infiltration by a leiomyosarcoma of the vaginal wall; cystectomy 2, multifocal pT3G3 tumors). The location of each sample was documented (bladder mapping). Urothelial cells were microdissected for LOH (chromosomes 9, 17p) and FISH analysis (CDKI2 (9p21), FACC (9q22), p53 (17p13.1), and centromeric probes for corresponding chromosome). Exons 5 to 9 of the p53 gene were sequenced in all tumor samples. No chromosomal alterations were detected in the cystectomy specimen without urothelial malignancies. The tumor-bearing bladder showed an increasing frequency of deletions with increasing malignancy of the investigated lesions. LOH analysis detected deletions only on chromosomes 9p and 17p. In contrast, FISH analysis revealed deletions of all investigated genes at chromosomes 9p, 9q, and 17p in all samples analyzed (preneoplastic and neoplastic tissue). An identical p53 mutation in codon 281 was found in all 7 analyzable tumor samples. Combination of molecular data with histopathologic bladder mapping suggested a monoclonal development of the multifocal lesions mostly via intraurothelial migration. Our data strengthen the results from recently published studies that patients with advanced urothelial carcinoma seem to have a monoclonal panurothelial disease in most cases. FISH showed a much higher sensitivity for detection of chromosomal losses than classical LOH analysis, especially in preneoplastic and small lesions. Combining 3 molecular approaches together with histopathologic organ mapping presents a valuable tool to determine the clonality status of multifocal bladder tumors. FAU - Denzinger, Stefan AU - Denzinger S AD - Department of Urology, University of Regensburg, Germany. FAU - Mohren, Kristin AU - Mohren K FAU - Knuechel, Ruth AU - Knuechel R FAU - Wild, Peter J AU - Wild PJ FAU - Burger, Maximilian AU - Burger M FAU - Wieland, Wolf F AU - Wieland WF FAU - Hartmann, Arndt AU - Hartmann A FAU - Stoehr, Robert AU - Stoehr R LA - eng PT - Journal Article DEP - 20051215 PL - United States TA - Hum Pathol JT - Human pathology JID - 9421547 SB - IM MH - Chromosomes, Human, Pair 17/genetics MH - Chromosomes, Human, Pair 9/genetics MH - Clone Cells/pathology MH - DNA Mutational Analysis MH - Female MH - Genes, p53/*genetics MH - Humans MH - In Situ Hybridization, Fluorescence/methods MH - *Loss of Heterozygosity MH - Male MH - Microsatellite Repeats MH - Middle Aged MH - Urinary Bladder Neoplasms/*genetics/*pathology EDAT- 2006/01/24 09:00 MHDA- 2006/02/25 09:00 CRDT- 2006/01/24 09:00 PHST- 2005/05/04 00:00 [received] PHST- 2005/10/21 00:00 [revised] PHST- 2005/10/21 00:00 [accepted] PHST- 2006/01/24 09:00 [pubmed] PHST- 2006/02/25 09:00 [medline] PHST- 2006/01/24 09:00 [entrez] AID - S0046-8177(05)00600-3 [pii] AID - 10.1016/j.humpath.2005.10.014 [doi] PST - ppublish SO - Hum Pathol. 2006 Feb;37(2):143-51. doi: 10.1016/j.humpath.2005.10.014. Epub 2005 Dec 15.