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Am J Physiol Cell Physiol 257: C743-C749, 1989;
0363-6143/89 $5.00
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AJP - Cell Physiology, Vol 257, Issue 4 C743-C749, Copyright © 1989 by American Physiological Society

ARTICLES

Enhanced erythropoietin secretion in hepatoblastoma cells in response to hypoxia

M. Ueno, I. Seferynska, B. Beckman, J. Brookins, J. Nakashima and J. W. Fisher
Department of Pharmacology, Tulane University School of Medicine, New Orleans, Louisiana 70112.

Erythropoietin (Ep) levels in spent culture media of a Hep G2 human hepatoblastoma cell line were measured by radioimmunoassay (RIA), fetal mouse liver erythroid colony formation (FMLC), and the exhypoxic polycythemic mouse assay (EHPCMA). The Hep G2 cells at high density produced approximately 700 mU/ml Ep when measured with the RIA. On the other hand, the Ep levels when assayed in EHPCMA and FMLC were 50 and 2,600 mU/ml, respectively. The bioactivity in FMLC was completely neutralized by an antibody to purified human recombinant Ep, indicating that the erythropoietic activity in the Hep G2 spent culture medium was immunologically equivalent to Ep. Ep levels in the medium from low-density Hep G2 cells in 5% O2 and 1% O2 were 2.5- and 4-fold greater, respectively, than that of 20% O2. In contrast, hyperoxia (40% O2) significantly inhibited Ep production. A significant increase in Ep secretion was also observed when the cells were incubated with cobaltous chloride (2 X 10(-6) -2.5 X 10(-4) M). Tunicamycin (0.5 micrograms/ml), which inhibits N-linked glycosylation, significantly reduced the enhancement of Ep secretion induced by hypoxia (1% O2) without affecting cell growth. Forskolin and cholera toxin, each of which increased the levels of cyclic AMP in the Hep G2 cells by 40-fold, produced a significant (P less than 0.05) further increase in Ep secretion in the presence of hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS)


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E. C. McGary, I. J. Rondon, and B. S. Beckman
Post-transcriptional Regulation of Erythropoietin mRNA Stability by Erythropoietin mRNA-binding Protein
J. Biol. Chem., March 28, 1997; 272(13): 8628 - 8634.
[Abstract] [Full Text] [PDF]


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