Cell cycle control and DNA damage response of conditionally immortalized urothelial cells

Bradley P. Dixon, Jeff Henry, Brian J. Siroky, Albert Chu, Pamela A. Groen, John Bissler

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: Children with complex urogenital anomalies often require bladder reconstruction. Gastrointestinal tissues used in bladder augmentations exhibit a greatly increased risk of malignancy, and the bladder microenvironment may play a role in this carcinogenesis. Investigating the influences of the bladder microenvironment on gastrointestinal and urothelial cell cycle checkpoint activation and DNA damage response has been limited by the lack of an appropriate welldifferentiated rothelial cell line system. Methodology/Principal Findings: To meet this need, we have developed a well-differentiated conditionally immortalized urothelial cell line by isolating it from the H-2Kb-tsA58 transgenic mouse. These cells express a thermosensitive SV40 large T antigen that can be deactivated by adjustment of cell culture conditions, allowing the cell line to regain normal control of the cell cycle. The isolated urothelial cell line demonstrates a polygonal, dome-shaped morphology, expresses cytokeratin 18, and exhibits well-developed tight junctions. Adaptation of the urothelial cell line to hyperosmolal culture conditions induces expression of both cytokeratin 20 and uroplakin II, markers of a superficial urothelial cell or "umbrella cell." This cell line can be maintained indefinitely in culture under permissive conditions but when cultured under non-permissive conditions, large T antigen expression is reduced substantially, leading to increased p53 activity and reduced cellular proliferation. Conclusions/Significance: This new model of urothelial cells, along with gastrointestinal cell lines previously derived from the H-2Kb-tsA58 transgenic mouse, will be useful for studying the potential mechanisms of carcinogenesis of the augmented bladder. 2011 Dixon et al.

Original languageEnglish (US)
Article numbere16595
JournalPLoS One
Volume6
Issue number1
DOIs
StatePublished - Feb 7 2011
Externally publishedYes

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Cell Cycle Checkpoints
DNA damage
DNA Damage
cell cycle
Cells
cell lines
bladder
Cell Line
Urinary Bladder
DNA
cells
Cell culture
Viral Tumor Antigens
carcinogenesis
Transgenic Mice
Uroplakin II
Carcinogenesis
Keratin-20
genetically modified organisms
antigens

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Cell cycle control and DNA damage response of conditionally immortalized urothelial cells. / Dixon, Bradley P.; Henry, Jeff; Siroky, Brian J.; Chu, Albert; Groen, Pamela A.; Bissler, John.

In: PLoS One, Vol. 6, No. 1, e16595, 07.02.2011.

Research output: Contribution to journalArticle

Dixon, Bradley P. ; Henry, Jeff ; Siroky, Brian J. ; Chu, Albert ; Groen, Pamela A. ; Bissler, John. / Cell cycle control and DNA damage response of conditionally immortalized urothelial cells. In: PLoS One. 2011 ; Vol. 6, No. 1.
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