The effect of a three dimensional growth environment on cell death and stress protein expression

dc.contributor.advisorDiller, K. R. (Kenneth R.)en
dc.contributor.committeeMemberNajjar, Ameren
dc.contributor.committeeMemberDunn, Andrewen
dc.contributor.committeeMemberSuggs, Lauraen
dc.contributor.committeeMemberMerchant, Fatimaen
dc.creatorSong, Alfred Seunghoonen
dc.date.accessioned2012-07-02T21:01:20Zen
dc.date.available2012-07-02T21:01:20Zen
dc.date.issued2012-05en
dc.date.submittedMay 2012en
dc.date.updated2012-07-02T21:01:45Zen
dc.descriptiontexten
dc.description.abstractUnderstanding the cellular response thermal stress is important for improving thermoablative treatments of cancer. Cells generally respond to thermal stress by expressing heat shock proteins, or undergoing cell death by apoptosis or necrosis. Most of our detailed knowledge regarding these cellular phenomena has been gathered in vitro in two dimensional (2D) environments. Yet, little is known about how prostate cancer cells respond to thermal stress in a more physiologically relevant three dimensional (3D) environment. Several approaches were used to investigate this question, all of which focused on controlled heating of cells in both two dimensional (2D) and 3D culture. Tools and assays were developed to investigate cellular response to thermal stress in 2D and 3D environments. A computer-controlled heating apparatus was constructed to heat cell cultures to precise temperatures and durations. Three dimensional growth environments were produced using Matrigel, a commercially available extracellular matrix (ecm) mixture. Transcriptional expression of heat shock protein 70 (HSP70) was measured using a green fluorescent protein (GFP) reporter gene under the control of an HSP promoter. Apoptosis, necrosis and HSP70 transcription was measured using flow cytometry analysis. Quantitative polymerase chain reaction (qPCR) and microscopy revealed that transmembrane targets may be involved in the mechanism of the effect which 3D culture has on the cellular response to heat shock. The results herein demonstrate that the 3D growth environment, may be protective to the cell in that the percentage of cells that undergo apoptosis or necrosis when exposed to heat shock are reduced. Furthermore, HSP70 expression is enhanced in 3D culture at a specific thermal dose and integrins and heat shock proteins may be part of the mechanism by which the ecm exerts its protective effect against thermal stress.en
dc.description.departmentBiomedical Engineeringen
dc.format.mimetypeapplication/pdfen
dc.identifier.slug2152/ETD-UT-2012-05-5559en
dc.identifier.urihttp://hdl.handle.net/2152/ETD-UT-2012-05-5559en
dc.language.isoengen
dc.subject3D cultureen
dc.subjectThree dimensional cultureen
dc.subjectCell cultureen
dc.subjectHSPen
dc.subjectHSP70en
dc.subjectHeat shock proteinen
dc.subjectHeat shock protein 70en
dc.subjectThermal therapyen
dc.subjectThermoablativeen
dc.subjectAnoikisen
dc.subjectSynoikisen
dc.subjectApoptosisen
dc.subjectNecrosisen
dc.subjectProstate canceren
dc.subjectCanceren
dc.subjectHeat shocken
dc.subjectMinimally invasiveen
dc.subjectBioheat transferen
dc.titleThe effect of a three dimensional growth environment on cell death and stress protein expressionen
dc.type.genrethesisen
thesis.degree.departmentBiomedical Engineeringen
thesis.degree.disciplineBiomedical Engineeringen
thesis.degree.grantorUniversity of Texas at Austinen
thesis.degree.levelDoctoralen
thesis.degree.nameDoctor of Philosophyen

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