Estrogen response in the hFOB 1.19 human fetal osteoblastic cell line stably transfected with the human estrogen receptor gene

S. A. Harris, K. R. Tau, R. J. Enger, D. O. Toft, B. L. Riggs, T. C. Spelsberg

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

The gene coding for the human wild-type estrogen receptor (ER) was stably transfected into the human fetal osteoblastic cell line hFOB 1.19, a clonal cell line which is conditionally immortilized with a temperature sensitive mutant of SV40 large T antigen (tsA58). Five subclones were obtained which express various levels of ER mRNA and protein. The subclone with the highest level of functional (nuclear bound) ER, hFOB/ER9, contained 3,931 (±1,341) 17β-estradiol molecules bound/nucleus as determined by the nuclear binding (NB) assay. Using the dextran coated charcoal (DCC) method, the level of total cytosolic ER measured was 204 (±2) fmol/mg protein. This subclone was examined further for estradiol (E 2) responsiveness. The ER expressed in hFOB/ER9 cells was shown to be functional using a transiently transfected ERE-FK-luciferase construct. Expression of luciferase from this construct increased ~25-fold in hFOB/ER9 cells following 10 -9M E 2 treatment. This effect on ERE-TK-luciferase expression was both dose and steroid dependant. Further, treatment of hFOB/ER9 cells with 10 -9M E 2 resulted in a 2.5-4.0- fold increase in endogenous progesterone receptor (PR) levels detected by steroid binding assays, and a noticeable increase in both the A and B forms of PR by western blot assay. The establishment of this estrogen responsive human osteoblastic cell line should provide an excellent model system for the study of estrogen action on osteoblast function.

Original languageEnglish (US)
Pages (from-to)193-201
Number of pages9
JournalJournal of Cellular Biochemistry
Volume59
Issue number2
DOIs
StatePublished - 1995

Fingerprint

Estrogen Receptors
Estrogens
Genes
Luciferases
Cells
Cell Line
Assays
Estradiol
Progesterone Receptors
Steroids
Polyomavirus Transforming Antigens
Charcoal
Viral Tumor Antigens
Dextrans
Osteoblasts
Western Blotting
Proteins
Messenger RNA
Temperature
Molecules

Keywords

  • estrogen receptor
  • osteoblasts
  • stable transfection
  • SV40 large T antigen

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Estrogen response in the hFOB 1.19 human fetal osteoblastic cell line stably transfected with the human estrogen receptor gene. / Harris, S. A.; Tau, K. R.; Enger, R. J.; Toft, D. O.; Riggs, B. L.; Spelsberg, T. C.

In: Journal of Cellular Biochemistry, Vol. 59, No. 2, 1995, p. 193-201.

Research output: Contribution to journalArticle

Harris, S. A. ; Tau, K. R. ; Enger, R. J. ; Toft, D. O. ; Riggs, B. L. ; Spelsberg, T. C. / Estrogen response in the hFOB 1.19 human fetal osteoblastic cell line stably transfected with the human estrogen receptor gene. In: Journal of Cellular Biochemistry. 1995 ; Vol. 59, No. 2. pp. 193-201.
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