Abstract
Purpose: Colon cancer is a genetic disease, caused by mutations in different oncogenes and tumor-suppressor genes. The aim of this study is to evaluate the usefulness of real-time PCR SNP analysis as a new technique in the loss of heterozygosity (LOH) analysis at the E-cadherin gene locus in sporadic colon cancer. Methods: One-hundred cases of human sporadic colon cancer and corresponding normal tissue samples were analyzed using two flanking polymorphic markers commonly used in the LOH analysis at the E-cadherin gene locus by conventional VNTR–LOH analysis. Two intragenic E-cadherin SNP markers were analyzed using real-time PCR SNP analysis. Results: LOH (17.6%) was detected using flanking markers, however, no LOH was detected when the intragenic E-cadherin SNP markers were introduced into our study. Since these markers are intragenic they more accurately represent the status of the E-cadherin gene than the previously used flanking markers. Conclusion: In conclusion, real-time PCR SNP analysis was found to be more accurate, faster, simpler, and a more high-throughput method than the conventional VNTR–LOH analysis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bussemakers MJ, van Bokhoven A, Mees SG, Kemler R, Schalken JA (1993) Molecular cloning and characterization of the human E-cadherin cDNA. Mol Biol Rep 17:123–128
Cashion AK, Driscoll CJ, Sabek O (2004) Emerging genetic technologies in clinical and research settings. Biol Res Nurs 5:159–167
Cavallaro U, Schaffhauser B, Christofori G (2002) Cadherins and the tumour progression: is it all in a switch? Cancer Lett 176:123–128
Chen HC, Chu RY, Hsu PN, Hsu PI, Lu JY, Lai KH, Tseng HH, Chou NH, Huang MS, Tseng CJ, Hsiao M (2003) Loss of E-cadherin expression correlates with poor differentiation and invasion into adjacent organs in gastric adenocarcinomas. Cancer Lett 201:97–106
Debruyne P, Vermeulen S, Mareel (1999) The role of the E-cadherin/catenin complex in gastrointestinal cancer. Acta Gastroenterol Belg 62:393–402
Huiping C, Sigurgeirsdottir JR, Jonasson JG, Eiriksdottir G, Johannsdottir JT, Egilsson VV, Ingvarsson S (1999) Chromosome alterations and E-cadherin gene mutations in human lobular breast cancer. Br J Cancer 81:1103–1110
Huiping C, Kristjansdottir S, Jonasson JG, Magnusson J, Egilsson V, Ingvarsson S (2001) Alterations of E-cadherin and beta-catenin in gastric cancer. BMC Cancer 1:16–22
Ilyas M, Tomlinson IPM, Hanaby A, Talbot IC, Bodmer WF (1997) Allele loss, replication errors and loss of expression of E-cadherin in colorectal cancers. Gut 40:654–659
Kanazawa T, Watanabe T, Kazama S, Tada T, Koketsu S, Nagawa H (2002) Poorly differentiated adenocarcinoma and mucinous carcinoma of the colon and rectum show higher rates of loss of heterozygosity and loss of E-cadherin expression due to methylation of promoter region. Int J Cancer 102:225–229
Machado JC, Oliveira C, Carvalho R, Soares P, Berx G, Caldas C, Seruca R, Carniero F, Sobrinho-Simoes M (2001) E-cadherin gene (CDH1) promoter methylation as the second hit in sporadic diffuse gastric carcinoma. Oncogene 20:1525–1528
Moreno-Bueno G, Hardisson D, Sarrio D, Sanchez C, Cassia R, Prat J, Herman JG, Esteller M, Matias-Guiu X, Palacios J (2003) Abnormalities of E- and P-cadherin and catenin (beta-, gamma-catenin, and p120ctn) expression in endometrial cancer and endometrial atypical hyperplasia. J Pathol 199:471–478
Osman I, Scher H, Dalabagni G, Reuter V, Zhang ZF, Cordon-Corado C (1997) Chromosome 16 in primary prostate cancer: a microsatellite analysis. Int J Cancer 71:580–584
Pecina-Slaus N, Gall-Troselj K, Slaus M, Radic K, Nikuseva-Martic T, Pavelic K (2004) Genetic changes of the E-cadherin and APC tumour suppressor genes in clear cell renal cell carcinoma. Pathology 36:145–151
Sarrio D, Moreno-Bueno G, Hardisson D, Sanchez-Estevez C, Guo M, Herman JG, Gamallo C, Esteller M, Palacios J (2003) Epigenetic and genetic alterations of APC and CDH1 genes in lobular breast cancer: relationships with abnormal E-cadherin and catenin expression and microsatellite instability. Int J Cancer 106:208–215
Spaventi R, Pečur L, Pavelić K, Pavelić ZP, Spaventi Š, Stambrook PJ (1994) Human tumor bank in Croatia: a possible model for a small bank as a part of future European tumor bank network. Eur J Cancer 30:419
Tsuda H, Zhang WD, Shimosato Y, Yokota J, Terada M, Sugimura T (1990) Allele loss on chromosome 16 associated with progression of human hepatocellular carcinoma. Proc Natl Acad Sci USA 87:992–1005
Wijhoven BP, de Both NJ, van Dekken H, Tilanius HW, Dinjens WN (1999) E-cadherin gene mutations are rare in adenocarcinomas of the oesophagus. Br J Cancer 80:1652–1657
Acknowledgements
This work was supported in part by grants number 0098108 and 0098109, from the Ministry of Science and Technology, Republic of Croatia.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Čačev, T., Jokić, M., Spaventi, R. et al. Loss of heterozygosity testing using real-time PCR analysis of single nucleotide polymorphisms. J Cancer Res Clin Oncol 132, 200–204 (2006). https://doi.org/10.1007/s00432-005-0051-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-005-0051-y