Endocytosis of pro-cathepsin D into breast cancer cells is mostly independent of mannose-6-phosphate receptors

被引:1
作者
Laurent-Matha, V [1 ]
Farnoud, MR [1 ]
Lucas, A [1 ]
Rougeot, C [1 ]
Garcia, M [1 ]
Rochefort, H [1 ]
机构
[1] Univ Montpellier 1, INSERM, Unite Hormones & Canc, U148, F-34090 Montpellier, France
关键词
cathepsin D; mannose-6-phosphate IGFII receptor; endocytosis; breast cancer;
D O I
暂无
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Cathepsin D trafficking is altered in cancer cells, leading to increased secretion of the pro-enzyme, which can be reinternalized by the same cancer cells and by stromal cells. We studied pro-cathepsin D endocytosis in two human breast cancer cell lines (MDA-MB231, MCF-7) and in human normal fibroblasts. Pro-enzyme uptake was studied indirectly through immunofluorescence analysis of anti-pro-cathepsin D monoclonal antibodies internalized in living cells. Both cancer cell lines internalized the pro-cathepsin D-antibody complex into endosomal compartments in the presence of 10 mM mannose-6-phosphate, Non-malignant fibroblasts, which do not secrete pro-cathepsin D, only internalized anti-cathepsin D antibody when purified pro-cathepsin D was added and this endocytosis was totally inhibited by mannose-6-phosphate, Cathepsin D endocytosis in cancer cells was not mediated by lectins or another receptor binding the cathepsin profragment, It was not due to fluid endocytosis, since another protein pS2 secreted by MCF-7 was not endocytosed with its antibody in the same conditions. Double-immunofluorescence and confocal microscopy analyses revealed that antibodies specific to pro-cathepsin D (M2E8) and to the mannose-6-phosphate/IGFII receptor were co-internalized independently in non-permeabilized MDA-MB231 cells and MCF-7 cells, but not in fibroblasts, Moreover, when metabolically labelled pro-cathepsin D secreted by MCF-7 or MDA-MB231 cells was incubated with homologous or heterologous non-radioactive cells, the time-dependent uptake and maturation of the pro-enzyme into fib ro blasts were totally inhibited by mannose-6-phosphate, whereas they were not in the two breast cancer cell lines, The percentage of mannose-6-phosphate-independent binding of radioactively labelled pro-cathepsin D to MDA-MB231 cells at 16 degrees C was higher (7-8%) at low pro-cathepsin D concentration than at high concentration (1.5%), indicating the presence of saturable binding site(s) at the cell surface that are different from the mannose-6-phosphate receptors, We conclude that, in contrast to fibroblasts, breast cancer cells can endocytose the secreted pro-cathepsin D by a cell surface receptor that is different from the mannose-6-phosphate receptors or other lectins, The nature of this alternative receptor and its significance in the action of secreted pro-cathepsin D remain to be elucidated.
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收藏
页码:2539 / 2549
页数:11
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