Abstract
The detection of unknown mutations has proved complex and time consuming, and this is certainly the case for HNPCC. Germline mutations have been detected in five of the six human DNA mismatch repair genes (in hMLH1, hMSH2, hMSH6, hPMS1, hPMS2, but not hMSH3) in HNPCC patients (1-7) with hMLH1 and hMSH2 being the most frequently affected (8-9). Point mutations resulting in missense, nonsense and frameshift alterations are found in HNPCC, as well as mutations leading to splicing alterations (9). The detection of mutations in these genes has relied upon direct sequencing of genomic DNA. Scanning or prescreening methods have been investigated (12-15), but at this stage direct sequencing is the gold standard by which these methods are generally judged. One caveat however, is that this method does not detect deletions that span exons or entire genes, the frequency of which may be up to 6.5% of HNPCC cases (16). To be rigorous, it would be prudent to include direct sequencing along with Southern analysis. This chapter will focus on the direct sequencing of hMLH1 and hMSH2, although the overall strategy may also be used for the analysis of the other mismatch repair genes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Peltomaki, P. and de la Chapelle, A. (1997) Mutations predisposing to hereditary nonpolyposis colorectal cancer. Adv. Cancer Res. 71, 93–119.
Moslein, G., Tester, D. J., Lindor, N. M., Honchel, R., Cunningham, J. M., French, A. J., et al. (1996) Micro satellite instability and mutation analysis of hMSH2 and hMLH1 in patients with sporadic, familial and hereditary colorectal cancer. Hum. Mol. Genet. 5, 1245–1252.
Papadopoulos, N., Nicolaides, N. C., Wei, Y. F., Ruben, S. M., Carter, K. C., Rosen, C. A., et al. (1994) Mutation of a mutL homolog in hereditary colon cancer (see comments). Science 263, 1625–1629.
Nicolaides, N. C., Papadopoulos, N., Liu, B., Wei, Y. F., Carter, K. C., Ruben, S. M., et al. (1994) Mutations of two PMS homologues in hereditary nonpolyposis colon cancer. Nature 371, 75–80.
Liu, B., Parsons, R., Papadopoulos, N., Nicolaides, N. C., Lynch, H. T., Watson, P., et al. (1996) Analysis of mismatch repair genes in hereditary non-polyposis colorectal cancer patients (see comments). Nature Med. 2, 169–174.
Akiyama, Y., Sato, H., Yamada, T., Nagasaki, H., Tsuchiya, A., Abe, R., and Yuasa, Y. (1997) Germ-line mutation of the hMSH6/GTBP gene in an atypical hereditary nonpolyposis colorectal cancer kindred. Cancer Research, 57, 3920–3923.
Shin, K. H., Ku, J. L., and Park, J. G. (1999) Germline mutations in a polycytosine repeat of the hMSH6 gene in Korean hereditary nonpolyposis colorectal cancer. J. Hum. Genet. 44, 18–21.
Nystrom-Lahti, M., Wu, Y., Moisio, A. L., Hofstra, R. M., Osinga, J., Mecklin, J. P., et al. (1996) DNA mismatch repair gene mutations in 55 kindreds with verified or putative hereditary non-polyposis colorectal cancer. Hum. Mol. Genet. 5, 763–769.
Peltomaki, P. and Vasen, H. F. (1997) Mutations predisposing to hereditary nonpolyposis colorectal cancer: database and results of a collaborative study. The International Collaborative Group on Hereditary Nonpolyposis Colorectal Cancer. Gastroenterology 113, 1146–1158.
Buerstedde, J. M., Alday, P., Torhorst, J., Weber, W., Muller, H., and Scott, R. (1995) Detection of new mutations in six out of 10 Swiss HNPCC families by genomic sequencing of the hMSH2 and hMLH1 genes. J. Med. Genet. 32, 909–912.
Tannergard, P., Lipford, J. R., Kolodner, R., Frodin, J. E., Nordenskjold, M., and Lindblom, A. (1995) Mutation screening in the hMLH1 gene in Swedish hereditary nonpolyposis colon cancer families. Cancer Res. 55, 6092–6096.
Farrington, S. M., Lin-Goerke, J., Ling, J., Wang, Y., Burczak, J. D., Robbins, D. J., and Dunlop, M. G. (1998) Systematic analysis of hMSH2 and hMLH1 in young colon cancer patients and controls. Am. J. Hum. Genet., 63, 749–759.
Wu, Y., Nystrom-Lahti, M., Osinga, J., Looman, M. W., Peltomaki, P., Aaltonen, L. A., et al. (1997) MSH2 and MLH1 mutations in sporadic replication errorpositive colorectal carcinoma as assessed by two-dimensional DNA electrophoresis. Genes Chromosomes Cancer 18, 269–278.
Suspiro, A., Fidalgo, P., Cravo, M., Albuquerque, C., Ramalho, E., Leitao, C. N., and Costa, M. F. (1998) The Muir-Torre syndrome: a rare variant of hereditary nonpolyposis colorectal cancer associated with hMSH2 mutation. Am. J. Gastroenterol. 93, 1572–1574.
Borresen, A. L., Lothe, R. A., Meling, G. I., Lystad, S., Morrison, P., Lipford, J., et al. (1995) Somatic mutations in the hMSH2 gene in microsatellite unstable colorectal carcinomas. Hum. Mol. Genet. 4, 2065–2072.
Wijnen, J., Vanderklift, H., Vasen, H., Khan, P. M., Menko, F., Tops, C., et al. (1998) MSH2 genomic deletions are a frequent cause of HNPCC. Nature Genet. 20, 326–328.
Kolodner, R. D., Hall, N. R., Lipford, J., Kane, M. F., Rao, M. R., Morrison, P., et al. (1994) Structure of the human MSH2 locus and analysis of two Muir-Torre kindreds for msh2 mutations. Genomics 24, 516–526.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.
About this protocol
Cite this protocol
Cunningham, J.M., Tester, D.J., Thibodeau, S.N. (2001). Mutation Detection in Colorectal Cancers. In: M., S. (eds) Colorectal Cancer. Methods in Molecular Medicine, vol 50. Humana Press. https://doi.org/10.1385/1-59259-084-5:87
Download citation
DOI: https://doi.org/10.1385/1-59259-084-5:87
Publisher Name: Humana Press
Print ISBN: 978-0-89603-767-0
Online ISBN: 978-1-59259-084-1
eBook Packages: Springer Protocols