Understanding 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. / text