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Functional properties of a conditionally phenotypic, estrogen-responsive, human osteoblast cell line
P V Bodine1, J Green, H A Harris
1Women's Health Research Institute, Wyeth-Ayerst, Radnor, Pennsylvania 19087, USA. bodinep@war.wyeth.com
Journal of Cellular Biochemistry
|June 1, 1997
Summary
Researchers developed a new human osteoblast cell line, HOB-03-CE6, that expresses functional estrogen receptors (ERs). This cell line demonstrates key responses to estrogen, making it a valuable tool for studying estrogen
Area of Science:
- Bone Biology
- Endocrinology
- Cell Biology
Background:
- Osteoblasts are crucial cells in bone metabolism and are known targets of estrogen action.
- Understanding estrogen's effects on osteoblasts is vital for bone health and treating bone diseases.
- Existing in vitro models may not fully recapitulate the complex responses of human osteoblasts to estrogen.
Purpose of the Study:
- To identify and characterize a human osteoblast cell line expressing functional estrogen receptors (ERs).
- To evaluate the utility of this cell line as a model for studying estrogen's molecular mechanisms in bone.
- To investigate specific endogenous responses of osteoblasts to 17 beta-estradiol (17 beta-E2).
Main Methods:
- Screening of 66 conditionally immortalized human osteoblast cell lines for ER alpha mRNA and functional ER protein.
- Characterization of the selected cell line (HOB-03-CE6) for osteoblast-specific markers, differentiation, and response to various stimuli.
- Assessing estrogen receptor expression and function using quantitative RT-PCR, luciferase reporter assays, radioligand binding, and DNA gel-shift analysis.
- Evaluating 14 endogenous estrogen responses in HOB-03-CE6 cells.
Main Results:
- A clonal, conditionally immortalized human osteoblast cell line (HOB-03-CE6) was identified with functional ERs.
- The cell line expressed osteoblast markers, formed mineralized nodules, and responded to hormonal and growth factor stimuli.
- 17 beta-E2 upregulated luciferase activity in a dose-dependent manner, suppressed PTH-induced cAMP production and IL-6 mRNA, and upregulated alkaline phosphatase mRNA.
- Specific binding of [125I]-17 beta-E2 and ERE-binding activity in nuclear extracts confirmed functional ERs.
Conclusions:
- The HOB-03-CE6 cell line represents a valuable in vitro model for studying estrogen action in human osteoblasts.
- This model allows for the exploration of molecular mechanisms underlying estrogen's effects on bone cells.
- The identified responses to 17 beta-E2 provide insights into estrogen's role in regulating specific osteoblast functions.
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