Molecular Determination of the Human Pappillomavirus-18 Integration in the HeLa Cervical Cancer Cell Line
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Authors
Bachman, Jonathan
Issue Date
2001
Type
Thesis
Language
en_US
Keywords
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Abstract
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.
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v, 25 p.
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