136-P

Abstract

The accumulation of damages in the DNA of the cells along the time, seems to play an important paper in the pathogenesis of cancer patients. DNA damage accumulates especially in mtDNA over time, and hOGG1 is an enzyme responsible for the repair of DNA oxidative damage either in nuclei and mitochondria. The Ser326Cys polymorphism was shown to diminish the hOGG1 activity in vitro, and it was suggested that the 326Cys allele might pose a higher risk of 8-OHdG formation in DNA. Recently, some authors have demonstrated that the capacity to repair oxidative DNA damage was significantly decreased in healthy human subjects bearing the hOGG1 Cys326Cys homozygous variant genotype, compared to wild type individuals. In the present, we intend to study the possible relation of this polymorphism with Bladder Cancer, BC. Bladder tissue from 22 cases of BC, and 90 controls, from peripheral blood lymphocytes; genomic DNA was extracted by means of the DNA direct II (Dynal) kit. Genotyping for the hOGG1 Ser326Cys polymorphism was performed by PCR-RFLP technique, briefly, a 234 bp product was amplified using the forward: 5′-CCC AAC CCC AGT GGA TTC TCA TTG C-3′ and the reverse: 5′-GGT GCC CCA TCT AGC CTT GCG GCC CTT-3′ primers. PCR conditions were 4 min at 94 °C and followed by 30 cycles of 30 s at 94 °C, 30 s at 60 °C, and 1 min and 30 s at 72 °C, and a final elongation step of 5 min at 72 °C. Overnight digestion at 37 °C with Fnu4HI resulted in 213- and 21-bp products for the wild type allele (Ser326), and in 164-, 49- and 21-bp products for the mutant allele (Cys326). Digestion products were visualized after electrophoresis on a 2% agarose gel containing ethidium bromide. The distribution of polymorphism in BC was: 64,3% SS, 33% SC 2,7% CC, in the control group: 66,4 SS, 30,4 SC 3,2 CC. The Chi-square test, p < 0.001.There are not correlation in polymorphism in this study inter BC and control group.