title: HPV integration detection in CaSki and SiHa using detection of integrated papillomavirus sequences and restriction-site PCR.
abstract: Human Papillomavirus (HPV) infection is the primary cause of cervical neoplasia. HPV DNA is integrated into the human genome in the majority of cervical cancers. The nature of integration may differ with integration incorporating a single copy of HPV or occurring in concatenated form. Our understanding of HPV tumorigenesis is largely based on studies using characterised cell lines with defined integration sites; these cell lines provide an invaluable standard for validation of diagnostic assays. Cell lines also further understanding of integration mechanisms in clinical samples. The objective of this study was to explore integration assays and to investigate integration events in cell lines where HPV is integrated in concatenated form. Restriction site PCR and detection of integrated papillomavirus sequences were performed on DNA from SiHa and CaSki. A novel integration site on Xq27.3 and HPV genome rearrangements were detected in CaSki DNA. However, where integration was previously detected by FISH in CaSki, and reported to be integrated in concatenated form, integration was not detected by DIPS or RS-PCR. The data presented illustrate that HPV copy number can hinder integration detection; this needs consideration when interpreting results from tests applied to clinical samples.
authors.affiliation: Institute of Cancer Research and Genetics, Section of Obstetrics and Gynecology, Main Building, Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom. Electronic address: raybouldre1@cf.ac.uk.
authors.forename: Rachel
authors.initials: R
authors.lastname: Raybould
authors.affiliation: Institute of Cancer Research and Genetics, Section of Obstetrics and Gynecology, Main Building, Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Alison
authors.initials: A
authors.lastname: Fiander
authors.affiliation: Institute of Infection and Immunity, Section of Medical Microbiology, Tenovus Building, Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Gavin W G
authors.initials: GW
authors.lastname: Wilkinson
authors.affiliation: Institute of Cancer Research and Genetics, Section of Obstetrics and Gynecology, Main Building, Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Sam
authors.initials: S
authors.lastname: Hibbitts
mesh_terms: D045744:Cell Line, Tumor; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D016662:Virus Integration
keywords: CaSki; Cervical cancer; Human papillomavirus; SiHa; Viral integration