title: Determination of human papillomavirus 16 physical status through E1/E6 and E2/E6 ratio analysis.
abstract: Human papillomavirus (HPV) 16 genome integration into the host chromosome is a crucial event during the life cycle of the virus and a major step towards carcinogenesis. The integration of HPV16 DNA promotes a constitutive high expression level of E6 and E7 oncoproteins, resulting in the extensive proliferation of the infected epithelial cells. In the present report the physical status of the HPV16 genome was studied, through determination of E1/E6 and E2/E6 DNA copy number ratios in 61 cervical samples of low- and high-grade malignancy and 8 cervical cancer samples, all of them associated with HPV16 infection. The selection of E1, E2 and E6 amplification target regions was performed according to the most prevalent deleted/disrupted sites of E1 and E2 genes. For this target selection we also considered the most conserved regions of E1, E2 and E6 genes among the same HPV16 isolates that were recently reported by our group. The analysis of HPV16 DNA form revealed a significant association among the mixed DNA forms in low-grade and high-grade malignancies, (χ(2), P<0.01). The comparative analysis of E1/E6 and E2/E6 in the same cervical samples provides an accurate picture of HPV16 DNA form and may reveal whether different HPV16 DNA integrants coexist in the same cervical sample or not. This study proposes that E1/E6 and E2/E6 ratios determine with accuracy the HPV16 DNA integration pattern and may predict multiple integration events in the examined sample, thus providing significant information about the progression of cervical dysplasia.
authors.affiliation: Microbiology-Virology Laboratory, Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Larissa, Greece.
authors.forename: Dimitris
authors.initials: D
authors.lastname: Tsakogiannis
authors.affiliation: Microbiology-Virology Laboratory, Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Larissa, Greece.
authors.forename: Zaharoula
authors.initials: Z
authors.lastname: Kyriakopoulou
authors.affiliation: Microbiology-Virology Laboratory, Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Larissa, Greece.
authors.forename: Irina Georgia Anna
authors.initials: IGA
authors.lastname: Ruether
authors.affiliation: Microbiology-Virology Laboratory, Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Larissa, Greece.
authors.forename: Grigoris D
authors.initials: GD
authors.lastname: Amoutzias
authors.affiliation: Microbiology-Virology Laboratory, Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Larissa, Greece.
authors.forename: Tilemachos G
authors.initials: TG
authors.lastname: Dimitriou
authors.affiliation: Microbiology-Virology Laboratory, Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Larissa, Greece.
authors.forename: Valentina
authors.initials: V
authors.lastname: Diamantidou
authors.affiliation: Clinical Biochemistry Department, General Hospital of Athens, Athens, Greece.
authors.forename: Constantin
authors.initials: C
authors.lastname: Kotsovassilis
authors.affiliation: Microbiology-Virology Laboratory, Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Larissa, Greece.
authors.forename: Panayotis
authors.initials: P
authors.lastname: Markoulatos
mesh_terms: D002584:Cervix Uteri / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D018628:Gene Dosage; D052162:Human papillomavirus 16 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D030361:Papillomavirus Infections / Q000821:virology; D012097:Repressor Proteins / Q000235:genetics*; D016662:Virus Integration
chemical_list: D004279:DNA, Viral; D004268:DNA-Binding Proteins; C093478:E1 protein, Human papillomavirus 16; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins; C076036:oncogene protein E2, Human papillomavirus type 1