Analysis of CYP1B1 polymorphisms in lung cancer patients using novel, quick and easy methods based on CAPS and ACRS-PCR techniques

Abstract

Within the sequence of the CYP1B1 gene, more than 50 polymorphisms, resulting from single nucleotide changes (SNPs), have been described. Part of them play an important role as specific genetic markers in the process of carcinogenesis and for therapeutic purposes. In this publication, we present methods we have developed that enable specific and unambiguous identification of four polymorphisms that result in amino acid changes: c. 142C > G, c. 355G > T, c. 1294C > G, and c. 1358A > G. Our studies are based on CAPS and ACRS-PCR techniques, therefore they require only basic laboratory equipment and low financial outlays. Utilizing the described methods allows for the reduction of research time, cost, and the minimization of errors. Their effectiveness and efficiency depend on the careful design of appropriate primers and the precise selection of suitable restriction enzymes. As a result, further confirmation by sequencing is not necessary. Using the developed method, we examined 63 patients diagnosed with lung cancer and observed a 1.5 to 2.1 times higher frequency of the analyzed single nucleotide polymorphisms compared to the frequency in the European population. No correlation was found between the occurrence of the tested polymorphisms and the histopathological diagnosis or stage of cancer advancement.