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Visualization of endogenous p53-mediated transcription in vivo using sodium iodide symporter

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Authors
Kim, Kwang Il; Chung, June-Key; Kang, Joo Hyun; Lee, Yong Jin; Shin, Jae Hoon; Oh, Hyun Jeong; Jeong, Jae Min; Lee, Dong Soo; Lee, Myung Chul
Issue Date
2005-01-27
Publisher
American Association for Cancer Research
Citation
Clin Cancer Res. 2005 Jan 1;11(1):123-8.
Keywords
AnimalsBiological TransportBlotting, WesternCell Line, TumorDNA, Complementary/metabolismDose-Response Relationship, DrugDoxorubicin/pharmacology*Gene Expression Regulation, NeoplasticGenes, ReporterGenetic VectorsGentamicins/pharmacologyHumansMaleMiceMice, Inbred BALB CMice, NudeNeoplasm TransplantationPerchloric Acid/pharmacologyPlasmids/metabolismPotassium Compounds/pharmacologyRadionuclide ImagingReverse Transcriptase Polymerase Chain ReactionSymporters/*metabolismTime Factors*Transcription, GeneticTranscriptional ActivationTransfectionTumor Suppressor Protein p53/*metabolismUp-Regulation
Abstract
PURPOSE: To develop a gamma camera imaging method for the determination of endogenous gene expression, we evaluated the expression of endogenous p53 gene using human sodium iodide symporter (hNIS) gene as reporter. EXPERIMENTAL DESIGN: We constructed cis-p53RE-hNIS reporter vector placed under control of an artificial enhancer (p53RE). Moreover, we transfected it into human hepatoma cell line SK-Hep1 by liposome. Geneticin was used for the selection of stable transfectant (SK-Hep1p53NIS). To evaluate the function of hNIS, the inhibition study was examined with 1 mmol/L potassium perchlorate. After treatment of Adriamycin with serial dose for 24 hours, we measured the uptake of 125I and did Western blot analysis to evaluate expression of p53 protein. Tumor xenografts were produced in nude mice by s.c. injection of SK-Hep1p53NIS cells. After 7 days, scintigraphic images of nude mice before and after Adriamycin treatment were obtained using [99mTc]-pertechnetate. RESULTS: In the SK-Hep1p53NIS cells, Adriamycin-treated cells accumulated up to three times higher than did nontreated cells. Potassium perchlorate inhibited completely the uptake of 125I. As Adriamycin dose increased, radioiodide uptake was significantly correlated with activated p53 as well as total p53 protein level. When Adriamycin (2 mg/kg) was treated in the same mice, a significantly higher uptake of [99mTc]-pertechnetate was observed in SK-Hep1p53NIS xenografts compared with nontreated xenografts (P < 0.05, unpaired t test). CONCLUSIONS: These results suggest that p53 expression level can be monitored by NIS gene expression using cis-p53RE-hNIS system in vitro and in vivo.
ISSN
1078-0432 (Print)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15671536

http://hdl.handle.net/10371/28352
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College of Medicine/School of Medicine (의과대학/대학원)Nuclear Medicine (핵의학전공)Journal Papers (저널논문_핵의학전공)
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