Expression of E6/E7 Transforming Genes is not Altered During Stratification and Differentiation of Two HPV -18 Transformed Cervical Carcinoma Cell Lines in an Organotypic Culture System
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Authors
Noud, Patrick H.
Issue Date
1996
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Human papillomaviruses (HPV) are important pathogens that cause
papillomas: skin tumors that are often benign, but are occasionally malignant
and life threatening. The study of HPV has been slow to date because culture
systems that support HPV replication have been lacking. HPV replication is
differentiation-dependent, and requires a culture system that approximates
epithelial differentiation in vivo. Recently, organotypic raft systems have
been described that support HPV replication. The focus of this study was to
organotypically culture epithelial cells and HPV-positive cell lines, and
achieve levels of differentiation adequate to support replication of HPV. This
study also assessed the effects of differentiation on HPV-gene expression in
two HPV -positive cell lines. To characterize the organotypic raft culture
system, differentiation-specific markers were identified and monitored by
SDS-P AGE and immunoblotting. In addition, Reverse Transcriptase-Polymerase
Chain Reaction (RT-PCR) techniques were used to follow HPV
gene expression in the two HPV-positive cell lines as a function of the
differentiation state of the cells. The differentiation that I observed with the
organotypic raft system suggests that it will be suitable for the study of HPV in
vitro. RT-PCR and immunoblotting proved to be useful tools for
characterizing cellular differentiation and expression of HPV genes in this
culture system. Transfection of HPV genomes into keratinocytes prepared for
growth in organotypic rafts will provide an ultimate measure of the system's
ability to support HPV replication.
Description
v, 44 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.