title: Gamma irradiation and targeted radionuclides enhance the expression of the noradrenaline transporter transgene controlled by the radio-inducible p21(WAF1/CIP1) promoter.
abstract: The use of radiation-inducible promoters to drive transgene expression offers the possibility of temporal and spatial regulation of gene activation. This study assessed the potential of one such promoter element, p21(WAF1/CIP1) (WAF1), to drive expression of the noradrenaline transporter (NAT) gene, which conveys sensitivity to radioiodinated meta-iodobenzylguanidine (MIBG). An expression vector containing NAT under the control of the radiation-inducible WAF1 promoter (pWAF/NAT) was produced. The non-NAT expressing cell lines UVW (glioma) and HCT116 (colorectal cancer) were transfected with this construct to assess radiation-controlled WAF1 activation of the NAT gene. Transfection of UVW and HCT cells with pWAF/NAT conferred upon them the ability to accumulate [(131)I]MIBG, which led to increased sensitivity to the radiopharmaceutical. Pretreatment of transfected cells with γ radiation or the radiopharmaceuticals [(123)I]MIBG or [(131)I]MIBG induced dose- and time-dependent increases in subsequent [(131)I]MIBG uptake and led to enhanced efficacy of [(131)I]MIBG-mediated cell kill. Gene therapy using WAF1-driven expression of NAT has the potential to expand the use of this therapeutic modality to tumors that lack a radio-targetable feature.
authors: Experimental Targeted Radiation Therapeutics Group, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom. anthony.mccluskey@strath.ac.uk
authors: Anthony G
authors: AG
authors: McCluskey
authors: Robert J
authors: RJ
authors: Mairs
authors: Annette
authors: A
authors: Sorensen
authors: Tracy
authors: T
authors: Robson
authors: Helen O
authors: HO
authors: McCarthy
authors: Sally L
authors: SL
authors: Pimlott
authors: John W
authors: JW
authors: Babich
authors: Sue
authors: S
authors: Champion
authors: Marie
authors: M
authors: Boyd
mesh_terms: D045744:Cell Line, Tumor; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics*; D005720:Gamma Rays; D006801:Humans; D050484:Norepinephrine Plasma Membrane Transport Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D019076:Transgenes
chemical_list: C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D050484:Norepinephrine Plasma Membrane Transport Proteins