Analysis of CYP1B1 Polymorphisms in Lung Cancer Patients Using Novel, Quick and Easy Methods Based on CAPS and ACRS-PCR Techniques

• Published in: International journal of molecular sciences 2024-06, Vol.25 (12), p.6676
• Main Authors: Dąbrowski, Adam, Nowicki, Maciej, Budzyńska, Aleksandra, Suchodolski, Jakub, Ogórek, Rafał, Chabowski, Mariusz, Przywara, Katarzyna
• Format: Article
• Language: English
• Subjects: Aged; Cancer patients; Care and treatment; Communication; Cytochrome; Cytochrome P-450 CYP1B1 - genetics; Development and progression; Enzymes; Equipment and supplies; Ethylenediaminetetraacetic acid; Female; Genes; Genetic aspects; Genetic markers; Health aspects; Humans; Hydrocarbons; Laboratories; Laboratory equipment; Lung cancer; Lung Neoplasms - genetics; Male; Methods; Middle Aged; Mutation; Patients; Polymerase Chain Reaction - methods; Polymorphism; Polymorphism, Single Nucleotide; Single nucleotide polymorphisms; Smoking; Statistical analysis; Steroids; Tobacco smoke
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25126676

Within the sequence of the gene, more than 50 polymorphisms, resulting from single-nucleotide polymorphisms (SNPs), have been described. Some 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 the 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 cleaved amplified polymorphic sequences (CAPSs) and artificially created restriction site (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 and 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.