type of oxidative DNA damage (3). The modified base 8-hydroxy-2Ј-deoxyguanosine (8-OHdG) is a mutagenic lesion Alma Boninsegna, Aldo Vecchione 2 and Achille Cittadini in vivo and in vitro, leading to G-T and A-C substitutions (4).
Giovanni XXIII Cancer Research Center, Institute of General Pathology andIt is therefore suggested as a sensitive biomarker for molecular (GC/MS), high-performance liquid chromatography with electrochemical detection (HPLC-EC), 32 P-post-labelling In this study, the 8-hydroxy-2Ј-deoxyguanosine (8-OHdG) (PPL) and immunoassays (8)(9)(10)(11)(12). Due to the detection method level was assessed in human cervical cells by an immunoused, discrepancies in the measurement of 8-OHdG may arise. peroxidase method and was related to the presence ofIn fact, the hydrolysis and derivatization procedures needed human papillomavirus (HPV) infection and precancerous for GC/MS can introduce artefacts leading to an overestimation lesions. After optimizing the immunohistochemical method of oxidative damage; whereas HPLC might underestimate 8-of detecting oxidative DNA damage in whole cells, we haveOHdG due to incomplete enzymatic digestion of DNA, and used this technique to estimate the oxidative damage in the production of hydroxyl radicals during PPL assay can cervical cells collected during a routine PAP test. The cause artefactual formation of 8-OHdG from dG (13). For analysis of variance (ANOVA) of the data from human all these reasons, an immunoperoxidase method, using the samples showed significant differences in the 8-OHdG monoclonal antibody 1F7, has recently been developed for 8-content among normal, low-grade and high-grade squamOHdG detection in single cells (14). This assay has shown ous intraepithelial lesion (SIL, HGSIL and LGSIL, respectsufficient sensitivity for oxidative damage detection in liver ively; P < 0.001). In the comparison of the three groups, tissues from aflatoxin B 1 (AFB 1 )-treated rats and oral mucosa statistically significant differences were detected between cells from smokers and non-smokers (15).
normal SIL and HGSIL (P < 0.001) and between LGSILIn this study, after optimizing the method for the evaluation and HGSIL (P ⍧ 0.003), whereas no statistically significant of oxidative DNA damage in human cells, we have used this difference was found between normal SIL and LGSIL technique to estimate the oxidative damage and its possible (P ⍧ 0.1). Grouping observations by HPV status, no involvement in the development of cervical dysplasia. significant difference was detected in 8-OHdG levels between HPV ⍣ and HPV -subjects (P ⍧ 0.8). The polytomous and proportional odds models, extensions of the logistic Materials and methods regression analysis, showed that the effect of 8-OHdG levels Human subjects in rising the risk of dysplasia was roughly constant through Thirty-six women undergoing a routine Papanicolaou smear were enrolled in SIL grades. In conclusion, the immunoperoxidase method, the study. After informed consent was obtained, cervical cells collected by applied to si...