title: Direct detection of the AR-E211 G > A gene polymorphism from blood and tissue samples without DNA isolation.
abstract: The pathogenesis of prostate cancer (CaP) involves alterations in a gene structure of the androgen receptor (AR). The single nucleotide polymorphism AR-E211 G > A localized in exon 1 of the AR gene (G1733A) was detected using direct polymerase chain reaction and restriction digestion (PCR-RFLP) method on blood and tissue samples without prior DNA isolation. We used blood samples of patients with a diagnosis of benign prostatic hyperplasia (BPH) or CaP. From monitored group of CaP patients were selected specimen in formalin-fixed paraffin-embedded tissue blocks with morphology of BPH and CaP. The main objective of our study was to develop a method based the direct PCR-RFLP analysis from blood and tissue without prior DNA isolation for faster genotyping analysis of a large number of samples. We found no statistically significant differences in allelic % of the AR-E211 G > A polymorphism between BPH and CaP patients (p ≤ 0.8462). Genotyping of the AR-E211 G > A variant in blood was not identical with tumor tissue genotyping analysis. Significant agreement between blood and tissue AR-E211 G > A polymorphism only in non-tumor tissue focus was confirmed. Although we analyzed a limited number of the tissue samples, we suppose that a presence of the minor allele A may be associated with cancer transformation-induced changes of the modified AR gene.
authors.affiliation: Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, University Palacky Olomouc, Hnevotinska 3, 775 15, Olomouc, Czech Republic.
authors.forename: Silvie
authors.initials: S
authors.lastname: Reptova
authors.forename: Katerina Smesny
authors.initials: KS
authors.lastname: Trtkova
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Kolar
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D001769:Blood; D001774:Blood Chemical Analysis / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000235:genetics; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011470:Prostatic Hyperplasia / Q000097:blood / Q000178:diet therapy / Q000235:genetics / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000097:blood / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism; D011944:Receptors, Androgen / Q000097:blood / Q000235:genetics* / Q000378:metabolism
chemical_list: C410798:AR protein, human; D011944:Receptors, Androgen; D004247:DNA