8-oxo-7,8-dihydroguanine level – the DNA oxidative stress marker - recognized by fluorescence image analysis in sporadic uterine adenocarcinomas in women

Abstract

Objectives: In the case of carcinogenesis in human endometrium no information exists on tissue concentration of 8-oxo-7,8-dihydroguanine, the DNA oxidative stress marker. This was the main reason to undertake the investigation of this DNA modification in human uterine estrogen-dependent tissue cancers. Material and Methods: In order to estimate the level of oxidative damage, 8-oxo-7,8-dihydroguanine was determined directly in cells of tissue microscope slides using OxyDNA Assay Kit, Fluorometric. Cells were investigated under confocal microscope. Images of individual cells were captured by computer-interfaced digital photography and analyzed for fluorescence intensities (continuous inverted 8-bit gray-scale = 0 [black]-255 [white]). Fluorescence scores were calculated for each of 13 normal endometrial samples and 31 uterine adenocarcinoma specimens. Finally, the level of the oxidative stress marker was also analyzed according to histological and clinical features of the neoplasms. Results: The obtained data revealed that: 8-oxo-7,8-dihydroguanine levels were higher in uterine adenocarcinomas than in normal endometrial samples (48,32 vs. 38,64; p<0,001); in contrast to normal endometrium there was no correlation between age and DNA oxidative modification content in uterine cancer; highest mean fluorescence intensity was recognized in G2 endometrial adenocarcinomas; level of 8-oxo-7,8-dihydroguanine does not depend on Body Mass Index (BMI) and cancer uterine wall infiltration or tumor FIGO stage. Conclusions: Our study indicates that accumulation of the oxidized DNA base may contribute to the development of endometrial neoplasia, however, oxidative DNA damage does not seem to increase with tumor progression.