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Biomedical subjects

F X Maquart

Publications and source records attributed to F X Maquart.

At least 19 recordsLinked to original sources

[Role of matrikins in melanoma progression].

Expression of melanoma invasiveness, ultimately leading to the formation of metastases, requires that cancer cells break through the successive skin barriers (dermo-epidermal junction, dermis) constituted of various extracellular matrix constituents. In order to facilitate their progression, melanoma cells express, in concert with stromal cells, a group of proteolytic systems which degrade this extracellular structures. However, proteolysis of basement membrane, collagen or elastic fibers can uncover cryptic sites or/and liberate matrix fragments whose properties appeared distinct from their intact macromolecule counterparts. Those fragments, called matrikines, are able to empede or to accelerate melanoma progression ex vivo and in vivo. Non-collagenous domains of basement membrane collagens, which behave like potent "matstatins", are seen as potential pharmacological agents in melanoma.

Collagen↗

Nerve growth factor displays stimulatory effects on human skin and lung fibroblasts, demonstrating a direct role for this factor in tissue repair.

Nerve growth factor (NGF) is a polypeptide which, in addition to its effect on nerve cells, is believed to play a role in inflammatory responses and in tissue repair. Because fibroblasts represent the main target and effector cells in these processes, to investigate whether NGF is involved in lung and skin tissue repair, we studied the effect of NGF on fibroblast migration, proliferation, collagen metabolism, modulation into myofibroblasts, and contraction of collagen gel. Both skin and lung fibroblasts were found to produce NGF and to express tyrosine kinase receptor (trkA) under basal conditions, whereas the low-affinity p75 receptor was expressed only after prolonged NGF exposure. NGF significantly induced skin and lung fibroblast migration in an in vitro model of wounded fibroblast and skin migration in Boyden chambers. Nevertheless NGF did not influence either skin or lung fibroblast proliferation, collagen production, or metalloproteinase production or activation. In contrast, culture of both lung and skin fibroblasts with NGF modulated their phenotype into myofibroblasts. Moreover, addition of NGF to both fibroblast types embedded in collagen gel increased their contraction. Fibrotic human lung or skin tissues displayed immunoreactivity for NGF, trkA, and p75. These data show a direct pro-fibrogenic effect of NGF on skin and lung fibroblasts and therefore indicate a role for NGF in tissue repair and fibrosis.

Actins↗

The tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+ stimulates matrix metalloproteinase-2 expression by fibroblast cultures.

Glycyl-histidyl-lysine-Cu2+ (GHK-Cu) is a tripeptide-copper complex known to be a potent wound healing agent. We previously showed its ability to stimulate in vitro and in vivo the synthesis of extracellular matrix components. The aim of this study was to determine the effects of GHK-Cu on MMP-2 synthesis by dermal fibroblasts in culture. We showed that GHK-Cu increased MMP-2 levels in conditioned media of cultured fibroblasts. This effect was reproduced by copper ions but not by the tripeptide GHK alone. This stimulation was accompanied by an increase of MMP-2 mRNA level. We also showed that GHK-Cu increased the secretion of the tissue inhibitors of metalloproteinases, TIMP-1 and TIMP-2. Taken together, our results underline that GHK-Cu is not only an activator of connective tissue production but also of the remodeling of the extracellular matrix. It is able to modulate MMP expression by acting directly on wound fibroblasts.

Animals↗

Human UDP-glucose dehydrogenase gene: complete cloning and transcription start mapping.

UDP-glucose dehydrogenase (GDH) is an unique pathway enzyme, which furnishes in vertebrates the UDP-glucuronic acid for numerous transferases, including those of glycosaminoglycan synthesis and xenobiotics elimination. Using long and accurate PCR approach and searching the 5' cDNA-end sequences in public databases, we have cloned the human GDH gene. The gene contains 12 exons and spans over 26 kb. The first and eighth introns were not reported for mouse ortholog. Primer extension analysis identified the transcription start site 165 bases upstream from the translation initiation site. Most of the exons were interrupted on codon phase 0, confirming the conserved ancestral structure of the gene reported on the cDNA level.

Animals↗

Human blood monocytes interact with type I collagen through alpha x beta 2 integrin (CD11c-CD18, gp150-95).

Human blood monocytes are attracted into connective tissues during early steps of inflammation and wound healing, and locally interact with resident cells and extracellular matrix proteins. We studied the effects of type I collagen on monocyte adhesion and superoxide anion production, using human monocytes elutriated from peripheral blood and type I collagen obtained from rat tail tendon. Both acid-soluble and pepsin-digested type I collagens promoted the adhesion of monocytes, whereas only acid-soluble collagen with intact telopeptides induced the production of superoxide. Adhesion and activation of monocytes on acid-soluble type I collagen depended on the presence of divalent cations. mAbs directed against integrin subunits CD11c and CD18 specifically inhibited adhesion and activation of monocytes on type I collagen. Protein membrane extracts obtained from monocytes were submitted to affinity chromatography on collagen I-Sepharose 4B, and analyzed by Western blotting using specific anti-integrin subunit Abs. In the case of both acid-soluble and pepsin-digested collagens, two bands were revealed with mAbs against CD11c and CD18 integrin subunits. Our results demonstrate that monocytes interact with type I collagen through CD11c-CD18 (alpha x beta 2) integrins, which promote their adhesion and activation. For monocyte activation, specific domains of the type I collagen telopeptides are necessary. Interactions between monocytes and collagen are most likely involved in the cascade of events that characterize the initial phases of inflammation.

Acids↗

A specific sequence of the noncollagenous domain of the alpha3(IV) chain of type IV collagen inhibits expression and activation of matrix metalloproteinases by tumor cells.

The invasive properties of melanoma cells correlate with the expression of matrix metalloproteinases (MMPs) and their physiological modulators (tissue inhibitors of metalloproteinase and membrane-type MMPs) and with that of the alphaVbeta3 integrin. We investigated the effect of anterior lens capsule type IV collagen and of the alpha3(IV) collagen chain on the invasive properties of various tumor cell lines (HT-144 melanoma cells, HT-1080 fibrosarcoma cells). We demonstrated that anterior lens capsule type IV collagen or specifically the synthetic peptide alpha3(IV) 185-203 inhibited both the migration of melanoma or fibrosarcoma cells as well as the activation of membrane-bound MMP-2 by decreasing the expressions of MT1-MMP and the beta3 integrin subunit.

Amino Acid Sequence↗

Modulation of sulfated glycosaminoglycan and small proteoglycan synthesis by the extracellular matrix.

Cell culture in collagen lattice is known to be a more physiological model than monolayer for studying the regulation of extracellular matrix protein deposition. The synthesis of sulfated glycosaminoglycans (GAG) and dermatan sulfate (DS) proteoglycans by 3 cell strains were studied in confluent monolayers grown on plastic surface, in comparison to fully retracted collagen lattices. Cells were labelled with 35S-sulfate, followed by GAG and proteoglycan analysis by cellulose acetate and SDS-polyacrylamide gel electrophoresis, respectively. The 3 cell strains contracted the lattice in a similar way. In monolayer cultures, the major part of GAG was secreted into culture medium whereas in lattice cultures of dermal fibroblasts and osteosarcoma MG-63 cells but not fibrosarcoma HT-1080 cells, a higher proportion of GAGs, including dermatan sulfate, was retained within the lattices. Small DS proteoglycans, decorin and biglycan, were detected in fibroblasts and MG-63 cultures. They were preferentially trapped within the collagen gel. In retracted lattices, decorin had a higher Mr than in monolayer. Biglycan was detected in monolayer and lattice cultures of MG-63 cells but in lattice cultures only in the case of fibroblasts. In this last case, an up regulation of biglycan mRNA steady state level and down regulation of decorin mRNA was observed, in comparison to monolayers, indicating that collagen can modulate the phenotypical expression of small proteoglycan genes.

Adult↗

Implication of interleukin-4 in wound healing.

Interleukin-4 (IL-4) is a pleiotropic cytokine that is able to activate connective tissue cells and stimulate the accumulation of extracellular matrix macromolecules. In this report, the expression of IL-4 in normal wound healing was studied by immunohistochemistry. The effects of exogenous IL-4 or IL-4 antisense oligonucleotides administration were also studied in mouse experimental wounds. IL-4 expression was detected in the lower dermis below the wound as early as Day 1 after wounding. IL-4 expression was maximal by Day 4, then decreased progressively, and completely disappeared by Day 21 after wounding. Topical administration of IL-4 on experimental wounds in mice significantly accelerated the rate of healing, whereas IL-4 antisense oligonucleotides significantly inhibited healing. These results demonstrate that IL-4 may be implicated in normal wound healing.

Animals↗

Activation of fibroblasts in collagen lattices by mast cell extract: a model of fibrosis.

BACKGROUND: Mast cells are resident connective tissue cells able to secrete numerous inflammatory mediators in response to tissue aggression and might be implicated in the fibrotic processes. OBJECTIVES: To study the effects of mast cell products on fibroblast activity in connective tissues. METHODS: Mast cell extract was prepared by sonication of pure mast cell preparations obtained by peritoneal lavage of rats and added to the culture medium of fibroblast-populated collagen lattices. RESULTS: Mast cell extract was able to decrease the contraction of the collagen lattices, to stimulate total protein and collagen synthesis, and to increase the expression and activation of gelatinase A/MMP-2. CONCLUSION: These data are consistent with the hypothesis that mast cells in connective tissue may be responsible for fibroblast activation at the early phases of tissue repair and fibrosis.

Adult↗

Expression of glycosaminoglycans and small proteoglycans in wounds: modulation by the tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu(2+).

Glycyl-histidyl-lysine-Cu(2+) is a tripeptide-copper complex previously shown to be an activator of wound healing. We have investigated the effects of glycyl-histidyl-lysine-Cu(2+) on the synthesis of glycosaminoglycans and small proteoglycans in a model of rat experimental wounds and in rat dermal fibroblast cultures. Repeated injections of glycyl-histidyl-lysine-Cu(2+) (2 mg per injection) stimulated the wound tissue production, as appreciated by dry weight and total protein measurements. This stimulation was accompanied by an increased production of type I collagen and glycosaminoglycans (assessed, respectively, by hydroxyproline and uronic acid contents of the chamber). Electrophoretic analysis of wound tissue glycosaminoglycans showed an accumulation of chondroitin sulfate and dermatan sulfate in control wound chambers, whereas the proportion of hyaluronic acid decreased with time. The accumulation of chondroitin sulfate and dermatan sulfate was enhanced by glycyl-histidyl-lysine-Cu(2+) treatment. The expression of two small proteoglycans of the dermis, decorin and biglycan, was analyzed by northern blot. The biglycan mRNA steady-state level in the chamber was maximal at day 12, whereas the decorin mRNA increased progressively until the end of the experiment (day 22). Glycyl-histidyl-lysine-Cu(2+) treatment increased the mRNA level of decorin and decreased those of biglycan. In dermal fibroblast cultures, the stimulation of decorin expression by glycyl-histidyl-lysine-Cu(2+) was also found. In contrast, biglycan expression was not modified. These results show that the expression of different proteoglycans in wound tissue are regulated in a different manner during wound healing. The glycyl-histidyl-lysine-Cu(2+) complex is able to modulate the expression of the extracellular matrix macromolecules differently during the wound repair process.

Animals↗

Involvement of the 92-kDa gelatinase (matrix metalloproteinase-9) in the ceramide-mediated inhibition of human keratinocyte growth.

Triggering the ceramide pathway by exogenous treatment with neutral sphingomyelinase (Smase) inhibited human keratinocyte growth rate, while having no influence on cell apoptosis. Increasing the ceramide content of keratinocytes with Smase (100 U/ml) or C6-ceramide (1 microM) enhanced matrix metalloproteinase (MMP)-9 production. On the contrary, levels of MMP-2 secretion were unchanged. The inhibition of keratinocyte growth rate induced by ceramide could be annihilated by a peptide hydroxamate MMP inhibitor or an MMP-9 blocking antibody. In addition, inhibiting MMP-9 activity in control keratinocyte culture was found to stimulate keratinocyte proliferation. These data suggest a pivotal function of MMP-9 in the control of keratinocyte growth.

Antibodies↗

Transforming growth factor beta-1 up-regulates clusterin synthesis in thyroid epithelial cells.

Porcine epithelial cells in primary culture seeded on plastic substratum form a monolayer containing pseudo-vesicles. When cultured in the presence of thyreotropin (TSH) thyrocytes adopted a follicular-like structure and formed clusters. Transforming growth factor beta-1 (TGFbeta1) induced a rapid spreading of the TSH-treated cells only. At the same time, TGFbeta1 enhanced clusterin protein and mRNA expression. The increase of clusterin synthesis was proportional to the TGFbeta1 concentration in the culture medium. Tunicamycin abolished the up-regulation of whole clusterin synthesis and morphological changes. The activator protein-1 binding site partly directed the TGFbeta1-stimulated clusterin expression. Phorbol ester caused rapid spreading of the cells with disappearance of vesicular and follicular structures. It decreased clusterin mRNA and protein expression, but increased Mr 45, 000 protein secretion in both TSH-treated and nontreated cells. Up-regulation of clusterin expression may be a marker of TGFbeta-mediated thyrocyte dedifferentiation.

Animals↗

Influence of oleic acid on the expression, activation and activity of gelatinase A produced by oncogene-transformed human bronchial epithelial cells.

Invasiveness and metastatic capacity of tumor cells have been related to increased expression and activation of gelatinase-A (MMP-2). 9-octadecenoic acid (oleic acid, OA), a long-chain cis-unsaturated fatty acid, has been shown to partially inhibit the formation of lung metastatic colonies in an ex vivo model of implantation of metastatic cells into nude mice. Reduction of metastasis formation was suggested to be due to a decrease of MMP-2 activity in tumor tissue extracts. Since regulation of MMP-2 activity occurs at different levels, including gene expression, pro-enzyme activation and finally active enzyme inhibition, we here investigated the precise level of the inhibitory effect of OA on MMP-2 activity by oncogene-transformed human bronchial epithelial cells (BZR cells). OA, at the dose of 5 x 10(-5) M, was shown to inhibit by 50% MMP-2 activity released from BZR cells. Northern-blot analysis of mRNA encoding MMP-2, MT1-MMP, the physiological activator of MMP-2, and TIMP-2, the natural inhibitor of MMP-2, revealed that OA did not alter the steady-state levels of MMP-2, MT1-MMP and TIMP-2 mRNA. Also, gelatin zymography demonstrated that the extent of MMP-2 activation was not modified by OA treatment. On the contrary, OA could inhibit the fluorogenic quenching substrate (7-methoxycoumarin-4-yl)acetyl-L-Pro-Leu-Gly-Leu-[N-3-(2,4-dinitrophenyl)-L-2, 3-diaminopropionyl]-Ala-Arg-NH2 (Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2) hydrolysis by recombinant MMP-2 with Ki = 4.3 microM. These data suggest that the beneficial influence of OA on the formation of lung metastatic colonies was independent of its influence on MMP-2 expression and/or activation, but could be attributed to inhibition of MMP-2 activity.

Animals↗

Biochemical alterations of uterine leiomyoma extracellular matrix in type IV Ehlers-Danlos syndrome.

Collagens and glycosaminoglycans were assessed in 6 leiomyomas from otherwise healthy ("control") patients and 1 leiomyoma from a patient with type IV Ehlers-Danlos syndrome. The type IV Ehlers-Danlos leiomyoma contained less type III collagen and dermatan sulfate than control leiomyomas. This might contribute to the formation of a defective collagen network in type IV Ehlers-Danlos syndrome.

Adult↗

Expression and activation of pro-gelatinase A by human melanoma cell lines with different tumorigenic potential.

The production of various proteolytic enzymes by tumor cells facilitate the invasion of solid tumors into surrounding tissues. We examined three cell lines (M1Dor, M4Be and M3Da) derived from malignant melanoma which exhibited different abilities to grow in nude mice following subcutaneous grafting. By in vitro invasion assay using Boyden-chambers technique, we found that none of those cell lines were able to invade the Matrigel. Several studies have substantiated the role of matrix metalloproteinases (MMP), mainly gelatinases MMP-9 and MMP-2, in melanoma cell invasion. Each cell line constitutively produced MMP-2 (but not MMP-9) in its latent form only, with stronger production for the most tumorigenic cell line in vivo (M3Da). Integrity of the MMP-2 activation process was studied since MMP-2 was also recovered as zymogen at the cell plasma membrane. All cell lines secreted TIMP-1 and TIMP-2 in a constitutive manner and again, but TIMP-2 production as well as MT1-MMP expression were found inversely related to their tumorigenic potential. Plating cells onto type I or type IV collagen did not trigger pro-MMP-2 activation; on the contrary, conversion of pro-MMP-2 to its active form could be evidenced when melanoma cell lines were seeded in a three dimensional type I collagen lattice.

Blotting, Northern↗