A human osteosarcoma cell line expressing herpes simplex type-1 thymidine kinase: studies with radiolabeled (E)-5-(2-iodovinyl)-2'-fluoro-2'-deoxyuridine.
BACKGROUND
(E)-5-(2-Iodovinyl)-2'-fluoro-2'-deoxyuridine (IVFRU) is a pyrimidine nucleoside analogue that accumulates selectively in murine cells expressing herpes simplex type-1 thymidine kinase (HSV-1 TK). The uptake of [(125)I]IVFRU in human 143B osteosarcoma cells transduced with a retroviral vector bearing the HSV-1 TK gene (143B-LTK cells) is now reported.


METHODS
HSV-1 TK gene expression in 143B-LTK cells was confirmed by Western blotting and reverse transcriptase (RT)-PCR. Cell and subcellular uptake of [(125)I]IVFRU was determined in cell culture, and whole body biodistribution after intravenous injection of [(125)I]IVFRU was determined using nude mice bearing implanted 143B or 143B-LTK tumors.


RESULTS
Although IVFRU was less toxic to the human cell line expressing HSV-1 TK (143B-LTK) than ganciclovir, both IVFRU and ganciclovir were not toxic to the cell line not expressing HSV-1 TK (143B). When cells were exposed to [(125)I]IVFRU in vitro, only the 143B-LTK cells accumulated radioactivity. The acid-soluble fraction from 143B-LTK cell lysates contained 8-fold greater activity than the acid-insoluble fraction after an 8-h exposure to [(125)I]IVFRU. Biodistribution of [(125)I]IVFRU in nude mice bearing subcutaneous 143B and 143B-LTK tumors revealed widespread distribution of the nucleoside in vivo but with specific localization in 143B-LTK tumors.


CONCLUSIONS
The underlying biochemical process of metabolic entrapment of IVFRU in human osteosarcoma cells expressing HSV-1 TK is responsible for selective localization in these cells. The differences in subcellular distribution into the nucleic acid fraction, and in cytotoxicity, reflect the importance of cell type and lineage as determinants of the performance of gene imaging radiopharmaceuticals.
Morin
Kevin W
KW
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Canada.
Duan
Weili
W
Knaus
Edward E
EE
McEwan
Alexander J B
AJ
Wiebe
Leonard I
LI
D001859:Bone Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D005467:Floxuridine / Q000031:analogs & derivatives* / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000201:enzymology*; D006801:Humans; D007457:Iodine Radioisotopes; D012516:Osteosarcoma / Q000378:metabolism*; D013937:Thymidine Kinase / Q000235:genetics*; D014018:Tissue Distribution
D007457:Iodine Radioisotopes; D005467:Floxuridine; C060868:5-(2-iodovinyl)-1-(2'-fluoro-2'-deoxyuridine); D013937:Thymidine Kinase