Ascorbic acid enhances iron-induced ferritin translation in human leukemia and hepatoma cells.
Ascorbate is an important cofactor in many cellular metabolic reactions and is intimately linked to iron homeostasis. Continuously cultured cells are ascorbate deficient due to the lability of the vitamin in solution and to the fact that daily supplementation of media with ascorbate is unusual. We found that ascorbate repletion alone did not alter ferritin synthesis. However, ascorbate-replete human hepatoma cells, Hep3B and HepG2, as well as K562 human leukemia cells achieved a substantially higher cellular ferritin content in response to a challenge with iron than did their ascorbate-deficient counterparts grown under standard culture conditions. Most of the elevation in ferritin content was due to an increase in de novo ferritin synthesis of greater than 50-fold, as shown by in vivo labeling with [35S]methionine and immunoprecipitation. RNA-blot analysis showed only minor changes in steady state levels of ferritin mRNA, suggesting that ascorbate enhances iron-induced ferritin synthesis primarily by post-transcriptional events. Transient gene expression experiments using chloramphenicol acetyltransferase reporter gene constructs showed that the ascorbate effect on ferritin translation is not mediated through the stem-loop near the translational start site that transduces ferritin synthesis in response to cytokines. The data suggest that ascorbate possibly modifies the action of the iron-responsive element on ferritin translation, although more precise structure-function studies are needed to clarify this issue. These data demonstrate a novel role of ascorbate as a signaling molecule in post-transcriptional gene regulation. The mechanism by which ascorbate modulates cellular iron metabolism is complex and requires additional detailed investigation.
Department of Medicine, Harvard Medical School, Brigham and Women's Hospital, Boston, Massachusetts 02115.
I
I
Toth
J T
JT
Rogers
J A
JA
McPhee
S M
SM
Elliott
S L
SL
Abramson
K R
KR
Bridges
D001205:Ascorbic Acid / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular; D005293:Ferritins / Q000235:genetics*; D006801:Humans; D007501:Iron / Q000502:physiology*; D007938:Leukemia; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D005293:Ferritins; D007501:Iron; D001205:Ascorbic Acid