Sucrose Density Gradient Analysis of 1, 25-Dihydroxyvitamin D3 Binding in Human Breast Tumors

Sylvia Christakos, Alan Sori, Stuart M. Greenstein, Thomas F. Murphy

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

The object of this investigation was to detect the type and number of 1, 25-dihydroxyvitamin D3 [1, 25-(OH)2D3]-binding proteins present in breast tumor tissue and thus to assess, by using sucrose density gradient analysis, the significance of the presence of 1, 25-(OH)2D3 binding in breast tumors. Cytosol fractions from 25 tumors (24 malignant breast tumors and 1 benign breast mass) were analyzed for the binding of vitamin D3 metabolites by sucrose density gradient ultracentrifugation. Cytosol fractions from 13 breast tumors exhibited binding of 1, 25-(OH)2D3 to a 4.1S macromolecule. Only 5 of the tumors (20%) showed a specific binding protein for 1, 25-(OH)2D3, which had a sedimentation coefficient of 3.2-3.4S, similar to that of the 1, 25-(OH)2D3 receptor protein found in intestinal cytosol. However, levels below 8 fmol/mg protein were observed for the positive malignant tumors compared to receptor concentrations of 286-563 fmol/mg protein which have been reported for the adult mammalian intestine. No correlation was found between the presence or absence of a specific 3.2-3.4S binding protein and the levels of estrogen receptors. Two of the breast tumors showed binding of 1, 25-(OH)2D3 to the 5-6S 25-hydroxyvitamin D3-binding protein. Cytosol, fractionated by precipitation with ammonium sulfate at 40% saturation in order to reduce interference from either the 5-6S binding protein or the 4.1S complex, showed no specific binding of 1, 25-(OH)2D3. Rachitic chick intestinal cytosol, vitamin D-deficient rat intestinal cytosol, and cytosol from MCF 7 cells, used as positive controls, consistently exhibited the characteristic 3.2-3.6S receptor. We conclude, due to a heterogeneous cellular population and possible serum contamination in human breast tumors, that the characterization of 1, 25-(OH)2D3 receptors in breast cancer cells compared to normal cells may provide more definitive conclusions concerning the relationship between 1, 25-(OH)2D3 receptors and malignancy.

Original languageEnglish (US)
Pages (from-to)686-691
Number of pages6
JournalJournal of Clinical Endocrinology and Metabolism
Volume56
Issue number4
DOIs
StatePublished - Apr 1983
Externally publishedYes

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

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