Zinc retention differs between primary and transformed cells in response to zinc deprivation.
Previous studies in our laboratory have demonstrated that reducing the availability of zinc with the extracellular metal chelator DTPA (diethylenetriaminepentaacetate) enhances, rather than inhibits, the thyroid hormone induction of growth hormone mRNA in GH3 rat anterior pituitary tumor cells. To understand the actions of the chelator on cellular zinc status, we observed the effects of DTPA on (65)Zn uptake and retention. DTPA reduced the uptake of (65)Zn by GH3 cells from the medium, but when GH3 cells were prelabeled with (65)Zn, it resulted in greater retention of the isotope. In primary hepatocytes, DTPA both reduced the uptake of (65)Zn from the medium and increased efflux from prelabeled cells. To investigate this difference, we studied the effects of DTPA on radioactive zinc flux in the H4IIE (rat hepatoma), MCF-7 (human breast cancer) and Hs578Bst (nontransformed human mammary) cell lines and in rat primary anterior pituitary cells. DTPA reduced the uptake of (65)Zn in all cell lines examined. DTPA increased the retention of (65)Zn in prelabeled H4IIE, MCF-7 and Hs578Bst cells but reduced it in primary pituitary cells. Time course experiments showed that (65)Zn efflux is shut down rapidly by DTPA in transformed cells, whereas the chelator causes greater efflux from primary hepatocytes over the first 6 h. Experiments with (14)C-labeled DTPA confirmed that this chelator does not cross cell membranes, showing that it operates entirely within the medium. Expression of ZnT-1, the efflux transporter, was not affected by DTPA in H4IIE cells. Thus, zinc deprivation enhanced zinc retention in established cell lines but increased efflux from primary cells, perhaps reflecting differing requirements for this mineral.
Dutta
Amrita
A
Department of Nutritional Sciences, University of Connecticut, CT 06269-4017, USA.
Schaller
Matthew
M
Franco
Aime T
AT
Sankavaram
Kavitha
K
Grattan
Bruce J
BJ
Freake
Hedley C
HC
D000818:Animals; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D002614:Chelating Agents / Q000493:pharmacokinetics / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008297:Male; D009928:Organ Specificity; D004369:Pentetic Acid / Q000493:pharmacokinetics / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D015032:Zinc / Q000172:deficiency* / Q000493:pharmacokinetics*; D015037:Zinc Radioisotopes / Q000493:pharmacokinetics
D027682:Cation Transport Proteins; D002614:Chelating Agents; D012333:RNA, Messenger; D015037:Zinc Radioisotopes; C092392:Slc30a1 protein, rat; D004369:Pentetic Acid; D015032:Zinc