Cell Cycle Control and DNA Damage Response of Conditionally Immortalized Urothelial Cells 英文参考文献.docVIP

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Cell Cycle Control and DNA Damage Response of Conditionally Immortalized Urothelial Cells 英文参考文献.doc

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Cell Cycle Control and DNA Damage Response of Conditionally Immortalized Urothelial Cells 英文参考文献

CellCycleControlandDNADamageResponseof ConditionallyImmortalizedUrothelialCells BradleyP.Dixon1,JeffHenry1,BrianJ.Siroky1,AlbertChu1,PamelaA.Groen2,JohnJ.Bissler1* 1DivisionofNephrologyHypertension,CincinnatiChildren’sHospitalMedicalCenter,Cincinnati,Ohio,UnitedStatesofAmerica,2DivisionofPathologyLaboratory Medicine,CincinnatiChildren’sHospitalMedicalCenter,Cincinnati,Ohio,UnitedStatesofAmerica Abstract Background: Children with complex urogenital anomalies often require bladder reconstruction. Gastrointestinal tissues usedinbladderaugmentationsexhibitagreatlyincreasedriskofmalignancy,andthebladdermicroenvironmentmayplay aroleinthiscarcinogenesis.Investigatingtheinfluencesofthebladdermicroenvironmentongastrointestinalandurothelial cell cycle checkpoint activation and DNA damage response has been limited by the lack of an appropriate well- differentiatedurothelialcelllinesystem. Methodology/PrincipalFindings:Tomeetthisneed,wehavedevelopedawell-differentiatedconditionallyimmortalized urothelialcelllinebyisolatingitfromtheH-2Kb-tsA58transgenicmouse.ThesecellsexpressathermosensitiveSV40largeT antigenthatcanbedeactivatedbyadjustmentofcellcultureconditions,allowingthecelllinetoregainnormalcontrolof thecellcycle.Theisolatedurothelialcelllinedemonstratesapolygonal,dome-shapedmorphology,expressescytokeratin 18, andexhibits well-developed tight junctions. Adaptation of the urothelial cellline tohyperosmolal culture conditions inducesexpressionofbothcytokeratin20anduroplakinII,markersofasuperficialurothelialcellor‘‘umbrellacell.’’Thiscell 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:Thisnewmodelofurothelialcells,alongwithgastrointestinalcelllinespreviouslyderivedfrom the H-2Kb-tsA58 transgenic mouse, will be useful for studying the potential mechanis