Beta-galactosidase as a marker of HSP70 promoter induction in Dunning R3327 prostate carcinoma cells.
Hyperthermia is known to improve the response of tumors to radiation or chemotherapeutic treatment when combined in multimodal strategies. The cellular response to hyperthermia is associated with the synthesis of heat shock proteins (HSP). To study the stress response in prostate cancer we have developed a clone of Dunning R3327 rat prostate carcinoma cells stably transfected with a gene construct containing the E. coli beta-galactosidase gene driven by the Drosophila HSP70 promoter. The measurement of beta-galactosidase serves as a rapid and semiquantitative assay of HSP70 gene activation. The Dunning cell clone showed evidence of incorporation of the HSP70/beta-galactosidase construct within the genomic DNA by Southern blot analysis. When compared to mock-transfected control cells, the clone showed minimal baseline beta-galactosidase activity, which significantly increased following a hyperthermic stress. The time course of beta-galactosidase elevation following heat stress paralleled the time course of cellular HSP70 elevation by Western blot analysis. These stably transfected Dunning R3327 cells may provide a useful tool to study the effects of hyperthermia, radiation, and chemotherapeutic agents on the cellular stress response and in the establishment of HSP70 as a marker of cellular resistance in the multimodal treatment of prostate cancer.
Roigas
J
J
Clinic of Urology, Charité Medical School, Humboldt University of Berlin, Germany.
Wallen
E S
ES
Loening
S A
SA
Moseley
P L
PL
D000230:Adenocarcinoma / Q000473:pathology*; D000818:Animals; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006979:Hyperthermia, Induced; D008297:Male; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011471:Prostatic Neoplasms / Q000473:pathology*; D051381:Rats; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase
D018840:HSP70 Heat-Shock Proteins; D001616:beta-Galactosidase