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Immunological detection and quantitation of alpha transforming growth factors in human breast carcinoma cells

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Summary

Alpha transforming growth factors (αTGFs) were immunologically detected in the concentrated conditioned medium (CM) prepared from four human breast cancer cell lines and from primary cultures of human mammary epithelial cells, and in the tissue extracts prepared from normal, benign, and malignant breast biopsies. Immunoreactive αTGFs were quantitated by a competitive radioimmunoassay (RIA) using affinity-purified polyclonal sheep anti-rat αTGF antibodies which react with human αTGF but not with human epidermal growth factor (EGF). The relative level of RIA-detectable αTGFs in the CM from the breast cancer cell lines MCF-7, ZR-75-1, T47-D, and MDA-MB-231, and from the CM of primary cultures of human mammary epithelial cells, ranged from 0.02 to 0.85 ng/ml. MCF-7 or ZR-75-1 cells grown in the presence of 17β-estradiol (10−8 M) for 48 h were found to release two- to three-fold more αTGFs into their CM than the same cells grown in the absence of estrogen. In detergent extracts prepared from normal breast tissue, a benign fibrocystic lesion, fibroadenomas and primary breast carcinomas, the relative αTGF concentrations were found to range from 1.5 to 6 ng/mg cell protein. No significant correlations were found between the αTGF levels and the pathological state of the tissues, the estrogen receptor status of the tumors, or the relative amounts of theras gene protein p21ras in the tissues as determined by Western immunoblot analysis. The question of biological relevancy of αTGF for human mammary tumors will require further studies on (a) synthesis and turnover of αTGF, (b) the relationship between immunoreactivity and biological activity of αTGF, and (c) differences in biological responsiveness of mammary tumor cells.

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Authors and Affiliations

  1. Laboratory of Tumor Immunology and Biology, National Cancer Institute, National Institutes of Health, Building 10, Room 8B07, 20892, Bethesda, MD, USA

    Isabelle Perroteau, David Salomon, Michele DeBortoli & William Kidwell

  2. Departments of Cell Biology and Physiology and Internal Medicine, University of Texas Health Science Center, 77225, Houston, TX, USA

    Parule Hazarika, Robert Pardue & John Dedman

  3. Department of Biochemistry, Rockefeller University, 1230 York Avenue, 10021, New York, NY, USA

    James Tam

Authors
  1. Isabelle Perroteau
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  2. David Salomon
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  3. Michele DeBortoli
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  4. William Kidwell
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  5. Parule Hazarika
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  6. Robert Pardue
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  7. John Dedman
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  8. James Tam
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Perroteau, I., Salomon, D., DeBortoli, M. et al. Immunological detection and quantitation of alpha transforming growth factors in human breast carcinoma cells. Breast Cancer Res Tr 7, 201–210 (1986). https://doi.org/10.1007/BF01806251

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