Increased MIBG uptake after transfer of the human norepinephrine transporter gene in rat hepatoma.
The transport of MIBG by the human norepinephrine transporter (hNET) seems to be the critical step in the treatment of MIBG-concentrating tumors. Therefore, we investigated whether the accumulation of MIBG may be induced by retroviral transfection of the hNET gene in Morris hepatoma cells.


METHODS
A bicistronic retroviral vector for the transfer of the hNET coding sequence and the hygromycin resistance gene was generated. Morris hepatoma cells (MH3924A) were infected with the respective retroviral particles, and hNET-expressing cell lines MHhNEThyg1 to MHhNEThyg9 were obtained through hygromycin selection. The uptake of (3)H-norepinephrine or (131)I-MIBG and the efflux of (131)I-MIBG were determined in transfected and wild-type cells. In addition, the (131)I-MIBG distribution was monitored in nude mice and rats bearing wild-type and hNET-expressing hepatomas.


RESULTS
hNET-expressing hepatoma cell lines accumulated up to 36 times more norepinephrine than did wild-type cells and 8 times more than did hNET-expressing neuroblastoma cell line SK-N-SH. The addition of nisoxetine, a selective inhibitor of noradrenaline uptake, inhibited norepinephrine uptake. Maximal (131)I-MIBG accumulation was observed 2 h after incubation and was followed by 43% efflux within 4 h after the (131)I-MIBG-containing medium had been removed. In vivo experiments performed with nude mice bearing both hNET-expressing and wild-type tumors showed a 10-fold-higher accumulation of (131)I-MIBG in transfected tumors than in wild-type tumors. The ex vivo calculations revealed doses of 605 and 75 mGy in hNET-expressing and wild-type tumor tissues, respectively.


CONCLUSIONS
Transduction of the hNET gene enables Morris hepatoma cells to accumulate norepinephrine and MIBG. However, the retention of MIBG is brief; therefore, the absorbed dose of radiation in vivo is not expected to be therapeutically effective.
Altmann
Annette
A
Clinical Cooperation Unit Nuclear Medicine, German Cancer Research Center, Heidelberg, Germany. A.Altmann@dkfz-heidelberg.de
Kissel
Maria
M
Zitzmann
Sabine
S
Kübler
Wolfgang
W
Mahmut
Miriam
M
Peschke
Peter
P
Haberkorn
Uwe
U
D019797:3-Iodobenzylguanidine / Q000493:pharmacokinetics*; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000000981:diagnostic imaging / Q000378:metabolism*; D003001:Cloning, Molecular; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000000981:diagnostic imaging / Q000378:metabolism*; D008297:Male; D008657:Metabolic Clearance Rate; D009368:Neoplasm Transplantation; D009638:Norepinephrine / Q000378:metabolism; D050484:Norepinephrine Plasma Membrane Transport Proteins; D009928:Organ Specificity; D011874:Radiometry; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012016:Reference Values; D027981:Symporters / Q000235:genetics / Q000378:metabolism*; D014018:Tissue Distribution; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured / Q000000981:diagnostic imaging / Q000378:metabolism
D050484:Norepinephrine Plasma Membrane Transport Proteins; D019275:Radiopharmaceuticals; D011994:Recombinant Proteins; C493227:SLC6A2 protein, human; C493226:Slc6a2 protein, rat; D027981:Symporters; D019797:3-Iodobenzylguanidine; D009638:Norepinephrine