F. Kebers

Inhibition of stromal matrix metalloproteases: Effects on breast-tumor promotion by fibroblasts

Co‐injection of fibroblasts with human epithelial breast‐tumor MCF7 cells in the presence of Matrigel enhances tumor growth in nude mice. While most of the matrix metalloproteinases (MMPs) have been shown to be produced by stromal cells, tumor cells such as MCF7 cells are unable to produce MMPs. We therefore, hypothesized that the tumor‐promoting effect of fibroblasts could be related to their production of MMPs. In order to inhibit stromal proteases, over‐production of TIMP‐2 was induced in MCF7 cells by in vitro retroviral‐mediated gene transfer. TIMP‐2‐producing MCF7 cells were then co‐injected with fibroblasts into nude mice. Alternatively, we evaluated the effect of Batimastat, a synthetic inhibitor of MMPs, on the tumorigenicity of MCF7 cells co‐inoculated with fibroblasts into nude mice. Both physiological (TIMP‐2) and synthetic (Batimastat) inhibitors of MMPs were able to abolish the tumor‐promoting effect of fibroblasts. On the contrary, they failed to modulate the tumorigenicity of MCF7 cells injected alone. Interestingly, Matrigel from which low‐molecular‐weight proteins or growth factors had been removed failed to favor the tumorigenicity of MCF7 cells inoculated with fibroblasts. These findings emphasize the importance of fibroblasts in cancer progression, and suggest that their role could be related at least in part to production of proteases which can induce the release of factors from the extracellular matrix. Int. J. Cancer 76:267–273, 1998.© 1998 Wiley‐Liss, Inc.

Demonstration in vivo that stromelysin-3 functions through its proteolytic activity.

Stromelysin-3 (ST3), a matrix metalloproteinase (MMP) expressed in aggressive carcinomas, has been shown to promote tumor development in different in vivo experimental models. However, the inability of its mature form to degrade extracellular matrix components casts doubt on whether ST3 functions in vivo as a protease. In this study, we evaluated whether the ST3 tumor-promoting effect could be ascribed to its proteolytic activity and whether this putative protease could be targeted with MMP inhibitors. Catalytically inactive mutant cDNA of human (h) ST3 or mouse (m) ST3 were generated and transfected into MCF7 cells. When injected into nude mice in the presence of matrigel, the mutant-bearing cells did not exhibit the enhanced tumorigenicity elicited by MCF7 cells transfected with wild-type ST3 cDNA. In a second approach, TIMP2 overproduction in MCF7 cells expressing hST3 was induced by retroviral infection. The co-expression of ST3 and TIMP2 failed to enhance the tumorigenicity of MCF7 cells. Notably, matrigel depleted of low-molecular-weight proteins and growth factors failed to promote the tumorigenicity of ST3-expressing MCF7 cells. These findings provide the first in vivo evidence that ST3 is indeed a protease that can modulate cancer progression by remodeling extracellular matrix and probably by inducing it to release the necessary microenvironmental factors. Thus, ST3 represents an interesting target for specific MMP inhibition.

Restricted expression of membrane type 1-matrix metalloproteinase by myofibroblasts adjacent to human breast cancer cells.

The membrane type‐1 matrix metalloproteinase (MT1‐MMP), a protease originally identified in breast carcinoma, is characterized by its capacity to activate other MMPs (MMP‐2 and MMP‐13) and to degrade extracellular matrix. Our study was undertaken to localize and identify the MT1‐MMP expressing cells in human breast adenocarcinomas. A textural analysis of images obtained by immunohistochemistry and in situ hybridization showed precisely the co‐expression of alpha smooth muscle actin (αSM actin) and MT1‐MMP in myofibroblasts. MT1‐MMP expression is confined to myofibroblasts in close contact with tumor cells. In sharp contrast, the expression of MMP‐2 was more widely distributed in both αSM actin positive and negative cells close to and at distance from cancer cell clusters. Our in vitro observations are consistent with the higher level of MT1‐MMP expression and of MMP‐2 activation observed in αSM actin positive fibroblasts derived from breast tumors, as compared to normal breast fibroblasts. Collectively, these results implicate myofibroblasts as major producer of MT1‐MMP in breast cancer and emphasize the importance of stromal‐epithelial cell interactions in their progression.

Cell—Cell and Cell—Matrix Interactions During Breast Cancer Progression

Cancer cells are not simply isolated islands of cells residing in a specific organ; they are surrounded by a modified extracellular matrix and by stromal cells of the host tissue, both of which influence tumor progression. The stroma peritumoral is different from the normal breast stroma and is critically involved in malignant growth.

Effects of a progestogen on normal human breast epithelial cell apoptosis in vitro and in vivo

Many investigators have reported cyclic proliferation of normal human breast epithelial cells. A delicate balance between proliferation and apoptosis (programmed cell death) ensures breast homeostasis. Both the follicular and luteal phases of the menstrual cycle are characterized by proliferation, whereas apoptosis occurs only at the end of the latter phase. In this study, we observed that the withdrawal of a synthetic progestin (nomegestrol acetate or NOMAC), but not continuous treatment with it, induced apoptosis of normal human breast epithelial cells in vitro and in women who applied NOMAC gel to their breasts. Furthermore, this apoptotic response was specific to normal breast cells, since withdrawal of NOMAC did not induce apoptosis of tumoral T47D cells in vitro or of fibroadenoma cells in women. These observations open up new perspectives in the prevention of hyperplasia and breast cancer.

Progesterone Receptor Activation. An Alternative to SERMs in Breast Cancer

Data regarding the effects of progesterone and a progestagen on human normal breast epithelial cell proliferation and apoptosis are presented here. In postmenopausal women, adding progesterone to percutaneously administrated oestradiol significantly reduces the proliferation induced by oestradiol. In vitro and in premenopausal women, stopping the administration of nomegestrol acetate triggers a peak of apoptosis. Fibro-adenoma and cancerous cells do not show this regulation of apoptosis. Progesterone seems to be important in normal breast homeostasis.