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

## TL;DR

This study introduces cost-effective methods to identify CYP1B1 gene polymorphisms in lung cancer patients, finding higher frequencies in this group compared to the general European population.

## Contribution

The paper presents novel, low-cost CAPS and ACRS-PCR methods for unambiguous identification of four CYP1B1 SNPs without the need for sequencing.

## Key findings

- The developed methods enable specific identification of four CYP1B1 polymorphisms with no need for sequencing.
- Lung cancer patients showed 1.5 to 2.1 times higher frequencies of the analyzed SNPs compared to the European population.
- The methods reduce research time, cost, and error rates using basic lab equipment and careful primer/enzyme design.

## Abstract

Within the sequence of the CYP1B1 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.