engleski

The application of real-time PCR SNP analysis of tumor suppressor genes in sporadic colon cancer

Colon cancer is a genetic disease, caused by inherited or acquired mutations in different oncogenes and tumor-suppressor genes. In the past decade, extensive efforts have been made to define the molecular mechanisms underlying development and progression of the colon cancer and to identify possible molecular targets for therapy. Molecular studies have shown that activating mutations of the Wnt signaling pathway are responsible for over 90 % of all colorectal cancers. E-cadherin is involved in control of intercellular adhesion and acts as an invasion suppressor. We examined 60 cases of human sporadic colon cancer and corresponding normal tissue samples to evaluate the loss of heterozygosity (LOH) at the APC and E-cadherin gene loci. DNAs were used for PCR, RFLP, VNTR, SNP, real-time PCR and LOH analysis. To analyze LOH at the APC gene loci we used three RFLP intragenic markers. For the LOH analysis of E-cadherin gene locus we used two VNTR extragenic polymorphic markers. We analyzed intragenic E-cadherin SNP marker by real-time PCR, as well. The SNP and real-time PCR were found to be faster, simpler and more high-throughput method than the conventional methods used in the previous analyses. We can conclude that using the SNP and real-time in the analysis of molecular changes in tumor suppressor genes involved in colon cancer adds to the speed and the informativity of this kind of studies.

real-time SNP PCR; E-cadherin; colon cancer

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