title: Adenoviral-mediated transfer of a heat-inducible double suicide gene into prostate carcinoma cells.
abstract: Tumor cells that express a fusion gene comprised of Escherichia coli cytosine deaminase (CD) and herpes simplex virus type 1 thymidine kinase (TK) sequences exhibit activation of and subsequent killing by the normally innocuous prodrugs 5-fluorocytosine and ganciclovir (Rogulski et al., Hum. Gene Ther., 8: 73-85, 1997). To target localized expression of this therapeutic gene, we have constructed a recombinant adenovirus containing the CD-TK fusion gene under the control of a human inducible heat shock protein 70 promotional sequence. Strong expression of the fusion gene product was induced by heating at 41 degrees C for 1 h. Expression levels obtained were dependent on the multiplicity of infection used and the incubation time after heat shock. Heat-induced expression of the CD-TK protein significantly reduced the survival of PC-3 cells in the presence of both 5-fluorocytosine and ganciclovir. These studies represent a novel form of gene therapy for the transduction and regulation of a double suicide gene in tumor cells and may provide a unique application for hyperthermia in cancer therapy.
authors.affiliation: Department of Radiation Oncology, William Beaumont Hospital, Royal Oak, Michigan 48073-6769, USA.
authors.forename: R V
authors.initials: RV
authors.lastname: Blackburn
authors.forename: S S
authors.initials: SS
authors.lastname: Galoforo
authors.forename: P M
authors.initials: PM
authors.lastname: Corry
authors.forename: Y J
authors.initials: YJ
authors.lastname: Lee
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000963:Antimetabolites / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D043525:Cytosine Deaminase; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology; D005437:Flucytosine / Q000494:pharmacology; D015774:Ganciclovir / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D018259:Herpesvirus 1, Human / Q000201:enzymology; D006358:Hot Temperature; D006801:Humans; D008297:Male; D009700:Nucleoside Deaminases / Q000096:biosynthesis / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000628:therapy*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism
chemical_list: D000963:Antimetabolites; D000998:Antiviral Agents; D011993:Recombinant Fusion Proteins; D005437:Flucytosine; D013937:Thymidine Kinase; D009700:Nucleoside Deaminases; D043525:Cytosine Deaminase; D015774:Ganciclovir