Abstract
Laser microdissection is an essential method for the investigation of the multistep carcinogenic process in the urinary bladder. Reliable detection of tumor-specific alterations which can be compromised by the presence of normal cells, requires microdissection of pure tumor cell populations (>80%) to detect loss of heterozygosity (LOH) by either fluorescence in situ hybridization (FISH) or sequence analysis. Multiple molecular methods need to be performed in the course of studying often-small lesions. This chapter describes in detail the use of laser microdissection, whole-genome amplification by improved primer extension preamplification (I-PEP)-polymerase chain reaction, and subsequent LOH, FISH, and sequencing analysis in the investigation of urothelial tumors and their precursor lesions. The combination of the described methods allows a wide spectrum of molecular investigations of tumor cells and helps to understand the fundamental alterations involved in urothelial carcinogenesis.
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References
Epstein, J. I., Amin, M. B., Reuter, V. R., and Mostofi, F. K. (1998) The World Health Organization/International Society of Urological Pathology consensus classification of urothelial (transitional cell) neoplasms of the urinary bladder. Bladder Consensus Conference Committee. Am. J. Surg. Pathol. 22, 1435–1448.
Filbeck, T., Pichlmeier, U., Knuechel, R., Wieland, W. F., and Roessler, W. (2002) Clinically relevant improvement of recurrence-free survival with 5-aminolevulinic acid induced fluorescence diagnosis in patients with superficial bladder tumors. J. Urol. 168,67–71.
Zaak, D., Kriegmair, M., Stepp, H., et al. (2001) Endoscopic detection of transitional cell carcinoma with 5-aminolevulinic acid: results of 1012 fluorescence endoscopies. Urology 57, 690–694.
Veltman, J. A., van Weert I, Aubele, M., et al. (2001) Specific steps in aneuploidization correlate with loss of heterozygosity of 9p21, 17p13 and 18q21 in the progression of pre-malignant laryngeal lesions. Int. J. Cancer 91,193–199.
Mostofi F. K, Davis C. J., and Sesterhenn, I. A. (1999) Histological typing of urinary bladder tumors. WHO, International Histological Classification of Tumors. Springer, New York.
Schlegel, J., Stumm, G., Scherthan, H., et al. (1995) Comparative genomic in situ hybridization of colon carcinomas with replication error. Cancer Res. 55,6002–6005.
Dietmaier, W., Hartmann, A., Wallinger, S., et al. (1999) Multiple mutation analyses in single tumor cells with improved whole genome amplification. Am. J. Pathol. 154, 83–95.
Stoehr, R., Knuechel, R., Boecker, J., et al. (2002) Histologic-genetic mapping by allele-specific PCR reveals intraurothelial spread of p53 mutant tumor clones. Lab Invest. 82, 1553–1561.
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© 2005 Humana Press Inc.
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Hartmann, A., Stoehr, R., Wild, P.J., Dietmaier, W., Knuechel, R. (2005). Microdissection for Detecting Genetic Aberrations in Early and Advanced Human Urinary Bladder Cancer. In: Murray, G.I., Curran, S. (eds) Laser Capture Microdissection. Methods in Molecular Biology™, vol 293. Humana Press. https://doi.org/10.1385/1-59259-853-6:079
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DOI: https://doi.org/10.1385/1-59259-853-6:079
Publisher Name: Humana Press
Print ISBN: 978-1-58829-260-5
Online ISBN: 978-1-59259-853-3
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