Regulated expression of erythropoietin by two human hepatoma cell lines
The development of a cell culture system that produces erythropoietin (Epo) in a regulated manner has been the focus of much effort. The authors have screened multiple renal and hepatic cell lines for either constitutive or regulated expression of Epo. Only the human hepatoma cell lines, Hep3B and HepG2, made significant amounts of Epo as measured both by radioimmunoassay and in vitro bioassay (as much as 330 milliunits per 10/sup 6/ cells in 24 hr). The constitutive production of Epo increased dramatically as a function of cell density in both cell lines. At cell densities < 3.3 x 10/sup 5/ cells per cm/sup 2/, there was little constitutive release of Epo in the medium. With Hep3B cells grown at low cell densities, a mean 18-fold increase in Epo expression was seen in response to hypoxia and a 6-fold increase was observed in response to incubation in medium containing 50 ..mu..M cobalt(II) chloride. At similar low cell densities, Epo production in HepG2 cells could be enhanced an average of about 3-fold by stimulation with either hypoxia or cobalt(II) chloride. Upon such stimulation, both cell lines demonstrated markedly elevated levels of Epo mRNA. Hence, both Hep3B and HepG2 cell lines provide an excellent in vitro system in which to study the physiological regulation of Epo expression.
- Research Organization:
- Howard Hughes Medical Institute, Boston, MA (USA)
- OSTI ID:
- 6879466
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:22
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ERYTHROPOIETIN
BIOCHEMISTRY
RADIOIMMUNOASSAY
CELL CULTURES
CYTOLOGICAL TECHNIQUES
HEPATOMAS
HYBRIDIZATION
IODINE 125
PHOSPHORUS 32
RNA
TUMOR CELLS
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DISEASES
ELECTRON CAPTURE RADIOISOTOPES
IMMUNOASSAY
IMMUNOLOGY
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MITOGENS
NEOPLASMS
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PROTEINS
RADIOASSAY
RADIOIMMUNOLOGY
RADIOISOTOPES
TRACER TECHNIQUES
550201* - Biochemistry- Tracer Techniques