engleski

Methylation status of the host cells gene promoters and human papillomavirus type 16 and 18 genes in cervical lesions

Promjena metilacije DNA je jedan od mogućih čimbenika odgovornih za zloćudne promjene povezane s infekcijom humanim papiloma virusom (HPV). Stoga je u ovom radu istražen stupanj metiliranosti promotora staničnih gena CCNA1, CDH1, C13ORF18, DAPK1, HIC1, RARβ2, hTERT1, hTERT2 i TWIST1 u uzorcima žena s različitim oštećenjima vrata maternice. Za utvrđivanje stanja metiliranosti promotora staničnih gena te gena L1 HPV-a tipa 18 u uzorcima s ovom infekcijom, primjenjena je metoda lančane reakcije polimerazom specifične za metilaciju (MSP). U uzorcima s infekcijom HPV-a tipa 16, broj i položaj metiliranih citozina unutar tri regije genoma HPV-a tipa 16 je utvrđen metodom bisulfitnog sekvenciranja. Najboljim metilacijskim biljezima predstadija raka naspram uzoraka sa citološki normalnim nalazom pokazali su se geni CCNA1 i C13ORF18, a biljezima karcinoma naspram predstadija, geni CCNA1, C13ORF18, RARβ2, hTERT1, hTERT2 i TWIST1. Metiliranost genoma HPV-a tipova 16 i 18 je bila uglavnom u skladu s dijagnozom i stupnjem metiliranosti staničnih gena.

cervical cancer; epigenetic changes

Change in DNA methylation level is probably one of the factors responsible for the malignant progression of cervical lesions caused by human papillomavirus (HPV). In this study, we investigated the methylation status of CCNA1, CDH1, C13ORF18, DAPK1, HIC1, RARβ2, hTERT1, hTERT2 and TWIST1 gene promoters in samples of women with different cervical lesions. To determine the methylation status of cells gene promoters and L1 gene of HPV 18 Methylation Specific PCR (MSP) was used, while the methylated cytosines in the three regions of HPV 16 genome were detected by bisulfite sequencing. The best methylation biomarkers for cervical lesions compared to normal cervices were CCNA1 and C13ORF18, while for cervical carcinoma compared to cervical lesions were CCNA1, C13ORF18, RARβ2, hTERT1, hTERT2 and TWIST1 genes. Methylation of HPV 16 and 18 genomes was in concordance with cervical diagnosis and methylation pattern of cellular genes.