Modulation of the Expression of Interstitial and Type-Iv Collagenases in Coculture of Ht1080 Fibrosarcoma Cells and Fibroblasts

in Invasion & Metastasis (1995), 15(5-6), 169-78

Members of the metalloproteinase family (MMPs) are known to play a crucial role in the metastatic cascade. Here, we report some investigations about the synthesis of interstitial and type-IV collagenases ... [more ▼]

Members of the metalloproteinase family (MMPs) are known to play a crucial role in the metastatic cascade. Here, we report some investigations about the synthesis of interstitial and type-IV collagenases (gelatinases A and B) in a model of coculture of human fibroblasts and HT 1080 fibrosarcoma cells. The interstitial collagenase activity, mainly found in the conditioned medium of fibroblasts, and its mRNA level were increased in the in vitro coculture model. In contrast, gelatinase A was produced by both cell types. The HT 1080 cells additionally synthesised gelatinase B. In coculture, an enhancement of gelatinase A and the presence of its activated form were observed. Northern blot analysis demonstrated that this enzymatic enhancement occurred at a pretranslational level. The stimulation of the interstitial collagenase activity was partially mediated through soluble factor(s), whereas increased gelatinase A appeared to require direct cell-cell interactions. The extracellular matrix component, type-I collagen, stimulated the enzymatic activities released by the individual cells, but it did not modulate the synthesis of interstitial collagenase in coculture. Our results demonstrate that distinct MMPs are modulated by distinct mechanisms, all depending on specific interactions between tumour cells and host fibroblasts. [less ▲]

Different Mechanisms of Extracellular Matrix Remodeling by Fibroblasts in Response to Human Mammary Neoplastic Cells

in Invasion & Metastasis (1993), 13(2), 72-81

Human breast tumors are often associated with a fibrotic reaction termed desmoplasia. Tumor cells may indirectly modulate the composition of the extracellular matrix by influencing fibroblast properties ... [more ▼]

Human breast tumors are often associated with a fibrotic reaction termed desmoplasia. Tumor cells may indirectly modulate the composition of the extracellular matrix by influencing fibroblast properties. They may also directly interact with collagen fibrils leading to retraction of the matrix. We have studied in vitro the influence of various human mammary tumor cells on the proliferation rate of normal human fibroblasts and on their level of collagen synthesis, as well as their release of collagenase activity. Interactions between neoplastic cells and collagen matrix were investigated by incorporation of tumor cells in collagen gels (lattices) and measurement of their retraction. All cells tested (HBL100, SW613, SA52, MDA-MB-231, MCF7, MCF7/6, MCF7 ras, BT20 and T47D) were able to modulate the composition of the extracellular matrix by one or several of the mechanisms investigated. Our results also demonstrate an opposite regulation of collagen and collagenase production. The effects on the collagen metabolism and on fibroblast proliferation are probably mediated by soluble cytokines since they are reproduced by incubating the fibroblasts in the presence of medium conditioned by tumor cells. The desmoplastic reaction may thus result from different mechanisms dependent upon tumor cell types. [less ▲]

The Stimulation of Fibroblasts' Collagen Synthesis by Neoplastic Cells Is Modulated by the Extracellular Matrix

in Matrix (Stuttgart, Germany) (1992), 12(3), 213-20

Human fibroblasts cocultured with neoplastic MCF7 cells produce increased amounts of collagen. A maximal stimulation requires direct cell-cell contacts between tumor cells and fibroblasts. However, this ... [more ▼]

Human fibroblasts cocultured with neoplastic MCF7 cells produce increased amounts of collagen. A maximal stimulation requires direct cell-cell contacts between tumor cells and fibroblasts. However, this effect could be reproduced, although to a lesser extent, by medium conditioned by MCF7 cells, suggesting that it is mediated by a factor produced by MCF7 cells and secreted, at least partly, under a soluble form (Noel et al., 1992). This Collagen Stimulating Factor ("COSF") present in the culture medium displayed a molecular mass between 3,500 to 10,000 daltons, bound to heparin and appeared to be different from the growth factors described until now. The "COSF" can be released from the surface of MCF7 cells by treatment with heparin. The aim of the present work was to investigate the influence of various extracellular matrix components on the production and the release of "COSF". A 3- to 4-fold enhancement of collagen synthesis was observed in coculture on plastic and collagen type I substrates without significant modification of the non-collagen proteins. The increased collagen synthesis was paralleled by an elevation of specific collagen mRNAs level suggesting a regulation at a pretranslational level. On the opposite, in the presence of soluble or insoluble laminin, this stimulation was abolished. Similarly, coculture on "reconstituted basement membrane matrix", matrigel, did not increase collagen production. The "COSF" was found to bind to matrigel and could be released from the basement membrane matrix by treatment with heparin. [less ▲]

A histologic study of the extracellular matrix during the development of glomerulosclerosis in murine chronic graft-versus-host disease.

in American Journal of Pathology (1992), 140(5), 1147-56

The development of glomerulosclerosis was studied in murine chronic graft-versus-host disease (GvHD), which is a model for human systemic lupus erythematosus. The authors investigated the distribution ... [more ▼]

The development of glomerulosclerosis was studied in murine chronic graft-versus-host disease (GvHD), which is a model for human systemic lupus erythematosus. The authors investigated the distribution patterns of six components of the extracellular matrix (ECM), i.e., laminin, fibronectin, collagen types I, III, IV, and VI during the course of the disease. All of these ECM components except collagen type I were found in the glomeruli of normal mice, where all of them were intrinsic constituents of the mesangium. Laminin, fibronectin, and collagen type IV were also found in the glomerular capillary walls. Starting 6 weeks after the induction of GvHD and continuing at week 8, the onset of an expansion of the mesangial matrix was observed. At the same time, the amounts of laminin, fibronectin, and collagen types IV and VI increased. Ten weeks after the onset of the disease, glomerulosclerosis developed. Traces of the interstitial collagen type I were found in sclerotic glomeruli. The levels of four ECM components, i.e., collagens III, IV, VI, and laminin were markedly decreased in the sclerotic glomeruli as compared with week 8. In contrast, the amount of fibronectin in the sclerotic glomeruli increased dramatically. Immunoelectron microscopic examination showed fibronectin in the sclerotic lesions, in contrast to laminin, collagen type I, and collagen type IV. It is concluded that the sclerotic lesions in murine chronic GvHD contain fibronectin. The small amounts of the ECM components laminin, as well as collagens III, IV, and VI in the sclerotic glomeruli in GvHD, might represent remnants of mesangial material and collapsed capillary walls. These components are probably replaced by increased production and/or accumulation of collagen type I and fibronectin. [less ▲]

Rat Chromosome 5 (Q22-23) Contains Elements That Control Cell Morphology and Interactions with the Extracellular Matrix: A Study of Normal Fibroblast X Malignant Hepatoma Cell Hybrids

in Experimental Cell Research (1991), 196(2), 164-71

Cell interactions with the extracellular matrix are consistently modified in neoplasia. Malignant transformation has been correlated with modifications in the synthesis and distribution of matrix ... [more ▼]

Cell interactions with the extracellular matrix are consistently modified in neoplasia. Malignant transformation has been correlated with modifications in the synthesis and distribution of matrix components and with alterations of cell adhesive properties to these components. A particular class of genes, able to suppress the transformed phenotype in normal cells, may be involved in those phenotypic changes. By studying somatic cell hybrids between mouse hepatoma (BWTG3) cells and normal rat skin fibroblasts (RSF), Islam and co-workers were able to localize a gene or a group of genes controlling anchorage dependence and cell growth in vitro. This (or these) gene(s) was (were) assigned to the q22-23 fragment of rat chromosome 5. In the present study, we compare the morphology and the interactions with the extracellular matrix proteins (laminin, fibronectin, and collagen IV) and the synthesis of these proteins by RSF X BWTG3 hybrid cells that had either retained (BS181p10) or lost (BS181a5) the q22-23 region of rat chromosome 5. Our results suggest that the rat 5q22-23 fragment controls a part of the cell differentiation program including morphology, attachment to extracellular matrix, and synthesis of some matrix proteins, particularly alpha 1 and alpha 2 chains of collagen IV. [less ▲]

Expression of Laminin by Human Fibroblasts, Ht1080 Fibrosarcoma Cells and Mcf-7 Breast Adenocarcinoma Cells. Lack of Regulation by the Cell Density and Extracellular Matrix

in Cell Biology International Reports (1991), 15(6), 499-509

We have cultured normal fibroblasts, fibrosarcoma HT1080 cells and breast adenocarcinoma MCF-7 cells on various substrates (plastic, collagen type I, laminin). All cell types used adhered on the three ... [more ▼]

We have cultured normal fibroblasts, fibrosarcoma HT1080 cells and breast adenocarcinoma MCF-7 cells on various substrates (plastic, collagen type I, laminin). All cell types used adhered on the three substrates with, however, a delayed attachment on laminin. On all substrates, cell grew as monolayer with the exception of MCF-7 cells that formed clusters on laminin. The epithelial MCF-7 cells as well as mesenchymal cells (fibroblasts and tumoral HT1080 cells) synthesized laminin and expressed mRNA coding for laminin B1 chain and for the 67 kD laminin binding protein. The levels of these mRNAs were not modulated by culture conditions which affect cell morphology nor by cell density. [less ▲]

Detection of Chloroplast DNA by Using Fluorescent Monoclonal Anti-Bromodeoxyuridine Antibody and Analysis of Its Fate During Zygote Formation in Chlamydomonas Reinhardtii

in Current Genetics (1990), 18(3), 259-63

A monoclonal anti-bromodeoxyuridine antibody conjugated to fluorescein was used to detect the chloroplast nucleoids after specific incorporation of bromodeoxyuridine (BUdR) into the chloroplast DNA of ... [more ▼]

A monoclonal anti-bromodeoxyuridine antibody conjugated to fluorescein was used to detect the chloroplast nucleoids after specific incorporation of bromodeoxyuridine (BUdR) into the chloroplast DNA of Chlamydomonas reinhardtii. The incorporation of BUdR was enhanced by simultaneous treatment with fluorodeoxyuridine (FUdR). The method was applied to analyze the fate of chloroplast DNA in zygotes resulting from mating between BUdR-treated gametes (mt+ or mt-) and untreated gametes of opposite mating-type. In crosses between wild-type strains, the nucleoids of mt+ origin remained in the large majority of zygotes whereas those of mt- origin most often disappeared within the first hours following copulation. In crosses of the type mat-3 mt+ x wild-type mt- (the mat-3 mutation permits a high transmission of chloroplast genes from the mt- parent), the nucleoids of mt- origin were generally not eliminated which indicates that the mat-3 mutation prevents the selective destruction of paternal chloroplast DNA in the zygote. [less ▲]

Reconstituted basement membrane matrix stimulates interstitial procollagenase synthesis by human fibroblasts in culture.

in Matrix (Stuttgart, Germany) (1990), 10(6), 373-7

Synthesis of interstitial collagenase by human fibroblasts was compared when cultured on plastic, in the presence or absence of soluble laminin, on a type I collagen gel and on a gel of basement membrane ... [more ▼]

Synthesis of interstitial collagenase by human fibroblasts was compared when cultured on plastic, in the presence or absence of soluble laminin, on a type I collagen gel and on a gel of basement membrane components (matrigel). Fibroblasts cultured on matrigel or on type I collagen gel displayed an increase in the steady-state levels of mRNA for interstitial procollagenase that was proportional to its enzymatic activity. Laminin, the main component of matrigel, had no effect on the interstitial collagenase synthesis by fibroblasts. We suggest that matrigel, which stimulates the interstitial collagenase production at a transcriptional step, could regulate the catabolic potential of fibroblasts. [less ▲]

Cooperativity between fibroblasts and melanoma cells during invasion

Conference (1989)