Molecular Determination of the Human Pappillomavirus-18 Integration in the HeLa Cervical Cancer Cell Line
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The development of cervical cancer is highly associated with human papillomavirus (HPV) infection. The integration of HPV into the genome of infected cervical cells is temporally associated with a malignant transformation. A relationship between the sites of HPV integration in cervical cancer and the position of the common fragile sites (CFSs) has been observed at the cytogenetic level. It has also been demonstrated that HPV -16 (one of the two most prevalent types of HPV found in cervical cancer) integrations in cervical tumors frequently occur within CFSs at the molecular level. In order to further explore this link, a PCR-based technique was used in order to isolate cellular sequences flanking the site of HPV -18 integration in an HPV infected cell line. A human BAC (bacterial artificial chromosome) clone was isolated based on these flanking sequences and used as a probe for Fluorescent in situ Hybridization (FISH) on metaphase chromosomes derived from peripheral blood lymphocytes cultured in the presence of aphidicolin and caffeine. The data from this project confirmed, at the molecular level, that the previously reported integration site of HPV -18 into the cervical carcinoma cell line HeLa is at chromosome 8q24.1 FISH analysis indicated that the integration is positioned distal, but in close proximity, to the common fragile site FRA8C.