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Cathepsin D in Breast Cancer Cells Can Digest Extracellular Matrix in Large Acidic Vesicles

Philippe Montcourrier, Paul H. Mangeat, Guillermo Salazar, Muriel Morisset, Alain Sahuquet and Henri Rochefort
Philippe Montcourrier
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Paul H. Mangeat
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Guillermo Salazar
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Muriel Morisset
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Alain Sahuquet
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Henri Rochefort
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DOI:  Published September 1990
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Abstract

In breast cancer cell lines, pro-cathepsin D is synthesized in excess and abnormally processed, resulting in its slower maturation and increased secretion into the culture medium. Since this lysosomal protease is only active at acidic pH, we have searched for acidic compartments other than lysosomes where cathepsin D might be active when MCF7 cells are plated on corneal extracellular matrix. We found large acidic intracellular vesicles (1.5 to 20 µm in diameter) by acridine organe and 3-(2,4-dinitroanilino)-3′-amino-N-methyldipropylamine staining, two fluorescent probes which reveal acidic compartments. These vesicles were actively acidified. They were 2- to 20-fold more abundant in MCF7 breast cancer cells and primary cultures of human breast cancers cells than in primary cultures of normal mammary epithelial cells. In living MCF7 cells, high resolution video-enhanced microscopy showed that these vesicles were mobile and intracellular. Double immunolocalization indicated that they contained mature cathepsin D (but no detectable pro-cathepsin D) and endocytosed extracellular material. This material (dextran, trans-ferrin, and extracellular matrix) and the association with other lysosomal enzymes varied according to the vesicles, suggesting their heterogeneity (large endosomes or phagosomes).

We conclude that, in breast cancer cells, cathepsin D may digest intracellularly phagocytosed and/or endocytosed extracellular matrix in large acidic vesicles. We propose that the higher expression of cathepsin D associated with the increased number of large acidic vesicles in breast cancer cells may facilitate digestion of basement membrane and consequently metastasis.

Footnotes

  • ↵1 This study was supported by the Institut National de la Santé et de la Recherche Médicale, the Centre National de la Recherche Scientifique, the Association pour la Recherche sur le Cancer, the University of Montpellier I, and the European Economic Community (Grant EEC 85 100001 BE02 PUJU 1).

  • ↵2 To whom requests for reprints should be addressed.

  • Received December 27, 1989.
  • Accepted June 19, 1990.
  • ©1990 American Association for Cancer Research.
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September 1990
Volume 50, Issue 18
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Cathepsin D in Breast Cancer Cells Can Digest Extracellular Matrix in Large Acidic Vesicles
Philippe Montcourrier, Paul H. Mangeat, Guillermo Salazar, Muriel Morisset, Alain Sahuquet and Henri Rochefort
Cancer Res September 15 1990 (50) (18) 6045-6054;

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Cathepsin D in Breast Cancer Cells Can Digest Extracellular Matrix in Large Acidic Vesicles
Philippe Montcourrier, Paul H. Mangeat, Guillermo Salazar, Muriel Morisset, Alain Sahuquet and Henri Rochefort
Cancer Res September 15 1990 (50) (18) 6045-6054;
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