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Cathepsins D, B, and L in transformed human breast epithelial cells

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Abstract

To investigate the regulation of lysosomal enzymes during carcinogenesis, we measured cathepsins (Cats) D, B, and L in MCF-10F, which is a human breast epithelial cell line, and cells evolved after treatment with carcinogen and transfected with c-Ha-ras oncogene. The clones used in this study, MCF-10FTras, D3, D3-1, and D3-1Tras, expressed no estrogen receptors and gradually increased invasive potential, while oncogenetransfected lines were also tumorigenic in SCID mice [16,19]. Cats D, B, and L were determined in the cells and in cell media using enzyme-linked immunosorbent assay (ELISA), specific enzyme activity measurements, and immunocytochemistry. The major intra- and extracellular lysosomal proteinase in these cells was Cat D (30–180 pm/mg), followed by Cat B (2–10 pm/mg) and Cat L (1–5 pm/mg). An inverse relationship between intracellular Cat D levels and invasive potential of carcinogen-treated and c-Ha-ras oncogene-transfected cell lines was observed. No significant changes in extracellular concentration of Cat D precursor in this series of cell lines was observed. Intracellular levels of Cats B and L were unchanged or slightly lower in carcinogentreated D3 and D3-1 cells, as well as in MCF-10FTras. On the other hand, in D3-1Tras cell line, evolving from c-Ha-ras transfected D3-1 line, 3.5 fold and 4.4 fold increases in Cat B and Cat L, respectively, but a 2 fold decrease in Cat D, were observed compared to the parental cell line. Immunocytochemical staining showed a granular, polarized perinuclear and cytoplasmic staining of cathepsins in all cell lines. Cysteine proteinases stained more frequently and more intensely in D3-1Tras compared to other lines, confirming the immunochemical assays. We hypothesize that several molecular events, caused by a carcinogen and an oncogene such as c-Ha-ras, are needed to increase Cat B and Cat L, but not Cat D, expression. Therefore, the cysteine and aspartic lysosomal proteinases are differentially expressed in the breast cell lines with more invasive phenotype.

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Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, Albert Einstein Medical Center, Philadelphia, Pennsylvania, USA

    Tamara T. Lah, Endre Kalman, Balasahib G. Shinde & Ierachmiel Daskal

  2. Department of Surgery, Albert Einstein Medical Center, Philadelphia, Pennsylvania, USA

    Robert Somers

  3. Department of Pathology, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA

    Gloria Calaf, Sandino Estrada, Enrique Salero & Jose Russo

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  1. Tamara T. Lah
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  2. Gloria Calaf
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  3. Endre Kalman
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  4. Balasahib G. Shinde
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  9. Ierachmiel Daskal
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Lah, T.T., Calaf, G., Kalman, E. et al. Cathepsins D, B, and L in transformed human breast epithelial cells. Breast Cancer Res Tr 39, 221–233 (1996). https://doi.org/10.1007/BF01806189

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