Estrogen binding and estrogenic responses in normal human osteoblast-like cells

David J. Benz, Mark R Haussler, Barry S. Komm

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

A finite human cell line was established from trabecular bone explants obtained from a 48-year-old woman. These cells, designated BG688, were characterized as osteoblast-like in phenotype using the following independent criteria: (1) the presence of histochemically detectable alkaline phosphatase (AP) activity; (2) response to the calciotropic hormone 1,25-(OH)2D3 as assessed by increased AP activity; (3) synthesis and secretion of the osteoblast-specific marker bone gla protein; and (4) expression of α1(I)-procollagen and α1(III)-procollagen mRNAs in a pattern similar to that of other osteoblast-like cell lines. In addition to these classic osteoblast markers, BG688 cells also possess approximately 2400 high-affinity (Kd = 0.45 nM) 17β-estradiol (E2) binding sites per cell. The binding of E2 to these sites is specific, and of the steroid hormone agonists tested, E2 and diethylstilbestrol elicited the greatest amount of competition with radiolabeled E2. BG688 cells were also shown to respond to a physiologic concentration (10 nM) of E2. In vitro translation products of poly(A)+ RNA obtained from control and hormone-treated cells revealed a pleiotropic influence of E2 on the relative abundance of several mRNAs as assessed by two-dimensional gel electrophoretic analysis of their corresponding peptides. E2 also elicits a twofold increase in the steady-state concentration of α1(I)-procollagen mRNA as demonstrated by northern blot hybridization. Thus, we here extend our previous data obtained in osteoblast-like osteosarcoma cells to indicate that a normal osteoblastic cell line is a target for the action of estrogen. The E2 responsiveness of these cells, coupled with their osteoblast-like phenotype, renders BG688 cells an attractive system in which to investigate the role of estrogens in the regulation of osteoblast function.

Original languageEnglish (US)
Pages (from-to)531-541
Number of pages11
JournalJournal of Bone and Mineral Research
Volume6
Issue number6
StatePublished - Jun 1991

Fingerprint

Osteoblasts
Estrogens
Procollagen
Messenger RNA
Hormones
Cell Line
Alkaline Phosphatase
Phenotype
Diethylstilbestrol
Osteocalcin
Osteosarcoma
Northern Blotting
Estradiol
Gels
Steroids
Binding Sites
Peptides

ASJC Scopus subject areas

  • Surgery

Cite this

Estrogen binding and estrogenic responses in normal human osteoblast-like cells. / Benz, David J.; Haussler, Mark R; Komm, Barry S.

In: Journal of Bone and Mineral Research, Vol. 6, No. 6, 06.1991, p. 531-541.

Research output: Contribution to journalArticle

@article{8da94f7825ad477e99584516f85b66a7,
title = "Estrogen binding and estrogenic responses in normal human osteoblast-like cells",
abstract = "A finite human cell line was established from trabecular bone explants obtained from a 48-year-old woman. These cells, designated BG688, were characterized as osteoblast-like in phenotype using the following independent criteria: (1) the presence of histochemically detectable alkaline phosphatase (AP) activity; (2) response to the calciotropic hormone 1,25-(OH)2D3 as assessed by increased AP activity; (3) synthesis and secretion of the osteoblast-specific marker bone gla protein; and (4) expression of α1(I)-procollagen and α1(III)-procollagen mRNAs in a pattern similar to that of other osteoblast-like cell lines. In addition to these classic osteoblast markers, BG688 cells also possess approximately 2400 high-affinity (Kd = 0.45 nM) 17β-estradiol (E2) binding sites per cell. The binding of E2 to these sites is specific, and of the steroid hormone agonists tested, E2 and diethylstilbestrol elicited the greatest amount of competition with radiolabeled E2. BG688 cells were also shown to respond to a physiologic concentration (10 nM) of E2. In vitro translation products of poly(A)+ RNA obtained from control and hormone-treated cells revealed a pleiotropic influence of E2 on the relative abundance of several mRNAs as assessed by two-dimensional gel electrophoretic analysis of their corresponding peptides. E2 also elicits a twofold increase in the steady-state concentration of α1(I)-procollagen mRNA as demonstrated by northern blot hybridization. Thus, we here extend our previous data obtained in osteoblast-like osteosarcoma cells to indicate that a normal osteoblastic cell line is a target for the action of estrogen. The E2 responsiveness of these cells, coupled with their osteoblast-like phenotype, renders BG688 cells an attractive system in which to investigate the role of estrogens in the regulation of osteoblast function.",
author = "Benz, {David J.} and Haussler, {Mark R} and Komm, {Barry S.}",
year = "1991",
month = "6",
language = "English (US)",
volume = "6",
pages = "531--541",
journal = "Journal of Bone and Mineral Research",
issn = "0884-0431",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Estrogen binding and estrogenic responses in normal human osteoblast-like cells

AU - Benz, David J.

AU - Haussler, Mark R

AU - Komm, Barry S.

PY - 1991/6

Y1 - 1991/6

N2 - A finite human cell line was established from trabecular bone explants obtained from a 48-year-old woman. These cells, designated BG688, were characterized as osteoblast-like in phenotype using the following independent criteria: (1) the presence of histochemically detectable alkaline phosphatase (AP) activity; (2) response to the calciotropic hormone 1,25-(OH)2D3 as assessed by increased AP activity; (3) synthesis and secretion of the osteoblast-specific marker bone gla protein; and (4) expression of α1(I)-procollagen and α1(III)-procollagen mRNAs in a pattern similar to that of other osteoblast-like cell lines. In addition to these classic osteoblast markers, BG688 cells also possess approximately 2400 high-affinity (Kd = 0.45 nM) 17β-estradiol (E2) binding sites per cell. The binding of E2 to these sites is specific, and of the steroid hormone agonists tested, E2 and diethylstilbestrol elicited the greatest amount of competition with radiolabeled E2. BG688 cells were also shown to respond to a physiologic concentration (10 nM) of E2. In vitro translation products of poly(A)+ RNA obtained from control and hormone-treated cells revealed a pleiotropic influence of E2 on the relative abundance of several mRNAs as assessed by two-dimensional gel electrophoretic analysis of their corresponding peptides. E2 also elicits a twofold increase in the steady-state concentration of α1(I)-procollagen mRNA as demonstrated by northern blot hybridization. Thus, we here extend our previous data obtained in osteoblast-like osteosarcoma cells to indicate that a normal osteoblastic cell line is a target for the action of estrogen. The E2 responsiveness of these cells, coupled with their osteoblast-like phenotype, renders BG688 cells an attractive system in which to investigate the role of estrogens in the regulation of osteoblast function.

AB - A finite human cell line was established from trabecular bone explants obtained from a 48-year-old woman. These cells, designated BG688, were characterized as osteoblast-like in phenotype using the following independent criteria: (1) the presence of histochemically detectable alkaline phosphatase (AP) activity; (2) response to the calciotropic hormone 1,25-(OH)2D3 as assessed by increased AP activity; (3) synthesis and secretion of the osteoblast-specific marker bone gla protein; and (4) expression of α1(I)-procollagen and α1(III)-procollagen mRNAs in a pattern similar to that of other osteoblast-like cell lines. In addition to these classic osteoblast markers, BG688 cells also possess approximately 2400 high-affinity (Kd = 0.45 nM) 17β-estradiol (E2) binding sites per cell. The binding of E2 to these sites is specific, and of the steroid hormone agonists tested, E2 and diethylstilbestrol elicited the greatest amount of competition with radiolabeled E2. BG688 cells were also shown to respond to a physiologic concentration (10 nM) of E2. In vitro translation products of poly(A)+ RNA obtained from control and hormone-treated cells revealed a pleiotropic influence of E2 on the relative abundance of several mRNAs as assessed by two-dimensional gel electrophoretic analysis of their corresponding peptides. E2 also elicits a twofold increase in the steady-state concentration of α1(I)-procollagen mRNA as demonstrated by northern blot hybridization. Thus, we here extend our previous data obtained in osteoblast-like osteosarcoma cells to indicate that a normal osteoblastic cell line is a target for the action of estrogen. The E2 responsiveness of these cells, coupled with their osteoblast-like phenotype, renders BG688 cells an attractive system in which to investigate the role of estrogens in the regulation of osteoblast function.

UR - http://www.scopus.com/inward/record.url?scp=0025784949&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025784949&partnerID=8YFLogxK

M3 - Article

C2 - 1887816

AN - SCOPUS:0025784949

VL - 6

SP - 531

EP - 541

JO - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

IS - 6

ER -