Expression and cellular localization of naturally occurring β estrogen receptors in uterine and mammary cell lines

Paula V Monje, Ricardo Boland

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The protein ER-α has been exhaustively characterized in estrogen-sensitive tissues and cell lines. However, little is known regarding the expression and cellular distribution of the newly identified ER-β protein. We first quantified the specific estradiol binding site content in the estrogen-responsive cell lines MCF-7 (mammary) and SHM (myometrial). In the two cell types, these sites were associated to the expression of both ER-α and -β isoforms. Native ER-β was visualized to reside inside the nucleus by means of conventional indirect immunofluorescence. The cells expressed ER-β as a tight ∼50 kDa triplet when resolved by sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE) and blotted using antibodies mapping different domains of the cloned ER-β version. When the cells were subjected to homogenization and differential centrifugation, a substantial proportion of ER-β immunolabeling was localized at membrane subfractions. ER-β expression and partitioning was confirmed by Ligand blotting assays using estrogen derivatives coupled to different macromolecular tags. However, ER-α was expressed as the major estrogen binding protein in both cell lines. Similar localization experiments were performed on HeLa cells (cervix). Though usually considered ER-negative, this cell line displayed basal significant estrogen binding capacity and co-expression of both ER isoforms. Taken as a whole, the results indicate that ER-β could be expressed as functional estrogen binding proteins among a dominant population of ER-α sites in the cell lines under study.

Original languageEnglish (US)
Pages (from-to)136-144
Number of pages9
JournalJournal of Cellular Biochemistry
Volume86
Issue number1
DOIs
StatePublished - Jan 1 2002
Externally publishedYes

Fingerprint

Estrogen Receptors
Estrogens
Breast
Cells
Cell Line
Carrier Proteins
Protein Isoforms
Centrifugation
Indirect Fluorescent Antibody Technique
HeLa Cells
Cervix Uteri
Sodium Dodecyl Sulfate
Estradiol
Assays
Proteins
Binding Sites
Tissue
Ligands
Derivatives
Membranes

Keywords

  • Estrogen receptor
  • HeLa
  • MCF-7
  • SHM
  • α and -β isoforms

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Expression and cellular localization of naturally occurring β estrogen receptors in uterine and mammary cell lines. / Monje, Paula V; Boland, Ricardo.

In: Journal of Cellular Biochemistry, Vol. 86, No. 1, 01.01.2002, p. 136-144.

Research output: Contribution to journalArticle

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