Cathepsin D in breast cancer cells can digest extracellular matrix in large acidic vesicles.
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 microns in diameter) by acridine orange 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, transferrin, 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.
Montcourrier
P
P
Unité Hormones et Cancer (U 148) INSERM, Montpellier, France.
Mangeat
P H
PH
Salazar
G
G
Morisset
M
M
Sahuquet
A
A
Rochefort
H
H
D001943:Breast Neoplasms / Q000201:enzymology*; D002402:Cathepsin D / Q000032:analysis / Q000502:physiology*; D004705:Endocytosis; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D006801:Humans; D014407:Tumor Cells, Cultured
D002402:Cathepsin D