42 results on '"Rigot V"'
Search Results
2. In vivo perimenstrual activation of progelatinase B (proMMP-9) in the human endometrium and its dependence on stromelysin 1 (MMP-3) ex vivo
- Author
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Rigot, V, Marbaix, Etienne, Lemoine, Pascale, Courtoy, Pierre J., Eeckhout, Yves, UCL - MD/MNOP - Département de morphologie normale et pathologique, and UCL - MD/BICL - Département de biochimie et de biologie cellulaire
- Subjects
Time Factors ,Blotting, Western ,Matrix Metalloproteinase Inhibitors ,Biochemistry ,Endometrium ,Mice ,Culture Techniques ,Animals ,Humans ,Enzyme Inhibitors ,Molecular Biology ,Menstrual Cycle ,Progesterone ,Enzyme Precursors ,Dose-Response Relationship, Drug ,Estradiol ,Antibodies, Monoclonal ,Metalloendopeptidases ,Cell Biology ,Enzyme Activation ,Gelatinases ,Female ,Matrix Metalloproteinase 3 ,Research Article ,Protein Binding - Abstract
Most matrix metalloproteinases (MMPs) are secreted as inactive proenzymes. Their expression is well documented in several human tissues, but their activators in vivo are still unknown. To address this question, the activation of progelatinase B (proMMP-9) in the human endometrium was selected as a model system. ProMMP-9 was detected by gelatin zymography in homogenates of fresh endometrial tissue sampled during all phases of the menstrual cycle, whereas its active form was observed only during the late secretory and menstrual phases. Furthermore, proMMP-9 was expressed and activated in endometrial explants sampled outside the perimenstrual phase and cultured in the absence of both progesterone and oestradiol, mimicking the menstrual condition in vivo. Analysis of such tissue cultures by gelatin zymography and Western blotting showed that activation of proMMP-9 depended on a secreted factor and was selectively inhibited by either a synthetic inhibitor of stromelysin 1 (MMP-3) or a monoclonal antibody that specifically blocks MMP-3, thus providing strong evidence for the activation of proMMP-9 in vivo by MMP-3. The activation of proMMP-3 was itself inhibited by a broad-range MMP inhibitor in most cultures, but seemed to involve multiple pathways, implying both serine proteinases and metalloproteinases, which could operate in parallel or sequentially.
- Published
- 2001
3. Migration properties of the human ovarian adenocarcinoma cell line IGROV1: Importance of ?v?3 integrins and vitronectin
- Author
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Carreiras, Franck, Rigot, V�ronique, Cruet, S�verine, Andr�, Fr�d�ric, Gauduchon, Pascal, Marvaldi, Jacques, Laboratoire de Biochimie Cellulaire - CNRS UPRESA 6032, Aix-Marseille Université - Faculté de pharmacie (AMU PHARM), and Aix Marseille Université (AMU)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
[SDV]Life Sciences [q-bio] ,ComputingMilieux_MISCELLANEOUS - Abstract
International audience
- Published
- 1999
4. P.314 Saccharomyces boulardii induit la migration des cellules épithéliales intestinales
- Author
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Canonici, A., primary, Siret, C., additional, Rigot, V., additional, and André, F., additional
- Published
- 2009
- Full Text
- View/download PDF
5. Temporal and spatial association of matrix metalloproteinases with focal endometrial breakdown and bleeding upon progestin-only contraception.
- Author
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UCL - MD/BICL - Département de biochimie et de biologie cellulaire, UCL - MD/MNOP - Département de morphologie normale et pathologique, Galant, Christine, Vekemans, M, Lemoine, Pascale, Kokorine, I., Twagirayezu, P, Henriet, Patrick, Picquet, C, Rigot, V, Eeckhout, Yves, Courtoy, Pierre J., Marbaix, Etienne, UCL - MD/BICL - Département de biochimie et de biologie cellulaire, UCL - MD/MNOP - Département de morphologie normale et pathologique, Galant, Christine, Vekemans, M, Lemoine, Pascale, Kokorine, I., Twagirayezu, P, Henriet, Patrick, Picquet, C, Rigot, V, Eeckhout, Yves, Courtoy, Pierre J., and Marbaix, Etienne
- Abstract
The pathogenesis of irregular endometrial bleeding, the main reason for stopping contraception with progestins only, is unknown. Based on the recent reappraisal of the mechanisms of menstrual bleeding, we hypothesized that matrix metalloproteinases initiate this disorder. Volunteers upon Norplant treatment provided endometrial biopsies at the start of a bleeding episode and during nonbleeding intervals. Focal stromal breakdown, collagen fiber lysis, and collagenase-1 messenger ribonucleic acid were evidenced in most bleeding endometria, but never in the nonbleeding ones. In the breaking down areas, immunolabeling for gelatinase A was strongly increased, and that of progesterone and estrogen receptors was decreased. Explants from bleeding endometria produced high collagenase and gelatinase activities, whereas release from nonbleeding endometria was negligible. Bleeding endometria released more latent and active forms of collagenase-1 and active gelatinases A and B, but less tissue inhibitor of metalloproteinases-1, than nonbleeding endometria. Collagenase-1 release closely correlated with that of interleukin-1alpha. In contrast, N:-acetyl-beta-hexosaminidase and tissue inhibitor of metalloproteinases-2 were similarly released in both groups. Thus, endometrial bleeding occurs together with focal stromal breakdown, collagen lysis, expression and activation of several matrix metalloproteinases, and decreased production of tissue inhibitor of metalloproteinases-1. These results may lead to new pharmacological treatment of this common medical problem.
- Published
- 2000
6. Circulating sex hormones and endometrial stromelysin-1 (matrix metalloproteinase-3) at the start of bleeding episodes in levonorgestrel-implant users.
- Author
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UCL - MD/MNOP - Département de morphologie normale et pathologique, UCL - MD/BICL - Département de biochimie et de biologie cellulaire, Marbaix, Etienne, Vekemans, M, Galant, Christine, Rigot, V, Lemoine, Pascale, Dubois, D., Picquet, C, Henriet, Patrick, Twagirayezu, P, Sufi, S, Eeckhout, Yves, Courtoy, Pierre J., UCL - MD/MNOP - Département de morphologie normale et pathologique, UCL - MD/BICL - Département de biochimie et de biologie cellulaire, Marbaix, Etienne, Vekemans, M, Galant, Christine, Rigot, V, Lemoine, Pascale, Dubois, D., Picquet, C, Henriet, Patrick, Twagirayezu, P, Sufi, S, Eeckhout, Yves, and Courtoy, Pierre J.
- Abstract
Unpredictable endometrial bleeding is the major side-effect of levonorgestrel-releasing s.c. implants (Norplant), otherwise a method of choice for long-term contraception. The mechanisms responsible for bleeding are still unknown and no reliable treatment is available. Several matrix metalloproteinases (MMP) are expressed and activated in human endometrium only at menstruation and specific synthetic inhibitors of MMP fully prevent the tissue breakdown that occurs in menstrual-like endometrial explants. To investigate whether MMP are inappropriately expressed and activated in Norplant-treated endometria during bleeding episodes, volunteers were recruited to provide blood and endometrial biopsies at the start of bleeding episodes and during non-bleeding intervals. Whereas serum concentrations of levonorgestrel and sex hormones showed no change at bleeding, except for a slight decrease of oestradiol concentration, the expression and activation of stromelysin-1 released by explants cultured for 1 day were consistently increased at the start of bleeding episodes. Furthermore, stromelysin-1 was immunolocalized in stromal cells within breakdown areas of several bleeding endometria, but not in non-bleeding endometria. These observations suggest that the expression and activation of stromelysin-1 participate in the initiation of bleeding episodes upon Norplant contraception. New strategies in the prevention and treatment of abnormal bleeding based on MMP control should be envisaged.
- Published
- 2000
7. La fonction de l’intégrine δ2ɛ1 est réprimée par des intégrines δv sur des cellules d’adénocarinome colique
- Author
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Defilles, C., primary, Delamarre, E., additional, Lissitzky, J.C., additional, Conesa, M., additional, Luis, J., additional, and Rigot, V., additional
- Published
- 2006
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8. C2-L’IGF-1 module les interactions entre l’intégrine δVɛ5 et le complexe cadhérine/caténines
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Gouranton, E, primary, Bryuneel, E, additional, Rigot, V, additional, Garrouste, F, additional, Remacle-Bonnet, M, additional, Pommier, G, additional, Marvaldi, J, additional, and André, F, additional
- Published
- 2006
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9. Circulating sex hormones and endometrial stromelysin-1 (matrix metalloproteinase-3) at the start of bleeding episodes in levonorgestrel-implant users
- Author
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Marbaix, E., primary, Vekemans, M., additional, Galant, C., additional, Rigot, V., additional, Lemoine, P., additional, Dubois, D., additional, Picquet, C., additional, Henriet, P., additional, Twagirayezu, P., additional, Sufi, S., additional, Eeckhout, Y., additional, and Courtoy, P.J., additional
- Published
- 2000
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10. Integrins and E-cadherin cooperate with IGF-I to induce migration of epithelial colonic cells
- Author
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Andr�, Fr�d�ric, primary, Rigot, V�ronique, additional, Thimonier, Jean, additional, Montixi, Christine, additional, Parat, Fabrice, additional, Pommier, Gilbert, additional, Marvaldi, Jacques, additional, and Luis, Jos�, additional
- Published
- 1999
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11. Integrin ligation and PKC activation are required for migration of colon carcinoma cells
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Rigot, V., primary, Lehmann, M., additional, Andre, F., additional, Daemi, N., additional, Marvaldi, J., additional, and Luis, J., additional
- Published
- 1998
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12. Chloride Channel Blockers Inhibit the Na+/I− Symporter in Thyroid Follicles in Culture
- Author
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Gerard, C., primary, Rigot, V., additional, and Penel, C., additional
- Published
- 1994
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13. Chloride Channel Blockers Inhibit the Na+/I−Symporter in Thyroid Follicles in Culture
- Author
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Gerard, C., Rigot, V., and Penel, C.
- Abstract
Porcine thyroid cells in culture are able to reorganize into well-polarized follicle-like structures in the presence of cAMP analogs. These follicles exhibit on their basolateral membrane domain the Na+/I−symporter which allows iodide to accumulate in the thyrocytes. The initial rate of iodide influx through the Na+/I−symporter is inhibited up to 98% by the chloride channel blockers. 5 nitro-2(3-phenylpropylamino)benzoic acid and 3′,5-Dichlorodiphenylamine-2-carboxylic acid are the most effective inhibitors, with a K0.5value of 60 μM. This inhibition is not secondary to inhibition of chloride transport. Other chloride transporter blockers have been studied but showed lesser activities.
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- 1994
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14. The effect of ibuprofen on accumulation of indium-111-labeled platelets and leukocytes in experimental myocardial infarction.
- Author
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Romson, J L, primary, Hook, B G, additional, Rigot, V H, additional, Schork, M A, additional, Swanson, D P, additional, and Lucchesi, B R, additional
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- 1982
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15. Deciphering the Crosstalk Between Myeloid-Derived Suppressor Cells and Regulatory T Cells in Pancreatic Ductal Adenocarcinoma.
- Author
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Siret C, Collignon A, Silvy F, Robert S, Cheyrol T, André P, Rigot V, Iovanna J, van de Pavert S, Lombardo D, Mas E, and Martirosyan A
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- Animals, Biomarkers, Carcinoma, Pancreatic Ductal pathology, Cell Line, Tumor, Humans, Immunomodulation, Immunophenotyping, Lymphocytes, Tumor-Infiltrating, Mice, Myeloid-Derived Suppressor Cells pathology, T-Lymphocytes, Regulatory pathology, Tumor Microenvironment genetics, Tumor Microenvironment immunology, Xenograft Model Antitumor Assays, Carcinoma, Pancreatic Ductal etiology, Carcinoma, Pancreatic Ductal metabolism, Cell Communication immunology, Myeloid-Derived Suppressor Cells immunology, Myeloid-Derived Suppressor Cells metabolism, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory metabolism
- Abstract
Pancreatic ductal adenocarcinoma (PDAC) is a fatal disease with rising incidence and a remarkable resistance to current therapies. The reasons for this therapeutic failure include the tumor's extensive infiltration by immunosuppressive cells such as myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs). By using light sheet fluorescent microscopy, we identified here direct interactions between these major immunoregulatory cells in PDAC. The in vivo depletion of MDSCs led to a significant reduction in Tregs in the pancreatic tumors. Through videomicroscopy and ex vivo functional assays we have shown that (i) MDSCs are able to induce Treg cells in a cell-cell dependent manner; (ii) Treg cells affect the survival and/or the proliferation of MDSCs. Furthermore, we have observed contacts between MDSCs and Treg cells at different stages of human cancer. Overall our findings suggest that interactions between MDSCs and Treg cells contribute to PDAC immunosuppressive environment., (Copyright © 2020 Siret, Collignon, Silvy, Robert, Cheyrol, André, Rigot, Iovanna, van de Pavert, Lombardo, Mas and Martirosyan.)
- Published
- 2020
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16. Structure-Based Virtual Screening Allows the Identification of Efficient Modulators of E-Cadherin-Mediated Cell-Cell Adhesion.
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Dalle Vedove A, Falchi F, Donini S, Dobric A, Germain S, Di Martino GP, Prosdocimi T, Vettraino C, Torretta A, Cavalli A, Rigot V, André F, and Parisini E
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- Antigens, CD genetics, Cadherins genetics, Crystallography, X-Ray, Humans, Molecular Docking Simulation, Molecular Dynamics Simulation, Neoplasm Invasiveness pathology, Protein Conformation, RNA Interference, RNA, Small Interfering genetics, Spheroids, Cellular, Tumor Cells, Cultured, Antigens, CD metabolism, Cadherins metabolism, Cell Adhesion physiology, Pancreatic Neoplasms pathology
- Abstract
Cadherins are a large family of transmembrane calcium-dependent cell adhesion proteins that orchestrate adherens junction formation and are crucially involved in tissue morphogenesis. Due to their important role in cancer development and metastasis, cadherins can be considered attractive targets for drug discovery. A recent crystal structure of the complex of a cadherin extracellular portion and a small molecule inhibitor allowed the identification of a druggable interface, thus providing a viable strategy for the design of cadherin dimerization modulators. Here, we report on a structure-based virtual screening approach that led to the identification of efficient and selective modulators of E-cadherin-mediated cell-cell adhesion. Of all the putative inhibitors that were identified and experimentally tested by cell adhesion assays using human pancreatic tumor BxPC-3 cells expressing both E-cadherin and P-cadherin, two compounds turned out to be effective in inhibiting stable cell-cell adhesion at micromolar concentrations. Moreover, at the same concentrations, one of them also showed anti-invasive properties in cell invasion assays. These results will allow further development of novel and selective cadherin-mediated cell-cell adhesion modulators for the treatment of a variety of cadherin-expressing solid tumors and for improving the efficiency of drug delivery across biological barriers.
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- 2019
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17. Dendritic cell-based vaccination: powerful resources of immature dendritic cells against pancreatic adenocarcinoma.
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Collignon A, Silvy F, Robert S, Trad M, Germain S, Nigri J, André F, Rigot V, Tomasini R, Bonnotte B, Lombardo D, Mas E, and Beraud E
- Abstract
Pancreatic adenocarcinoma (PAC) has a poor prognosis. One treatment approach, investigated here, is to reinforce antitumor immunity. Dendritic cells (DCs) are essential for the development and regulation of adaptive host immune responses against tumors. A major role for DCs may be as innate tumoricidal effector cells. We explored the efficacy of vaccination with immature (i)DCs, after selecting optimal conditions for generating immunostimulatory iDCs. We used two models, C57BL/6Jrj mice with ectopic tumors induced by the PAC cell line, Panc02, and genetically engineered (KIC) mice developing PAC. Therapeutic iDC-vaccination resulted in a significant reduction in tumor growth in C57BL/6Jrj mice and prolonged survival in KIC mice. Prophylactic iDC-vaccination prevented subcutaneous tumor development. These protective effects were long-lasting in Panc02-induced tumor development, but not in melanoma. iDC-vaccination impacted the immune status of the hosts by greatly increasing the percentage of CD8
+ T-cells, and natural killer (NK)1.1+ cells, that express granzyme B associated with Lamp-1 and IFN-γ. Efficacy of iDC-vaccination was CD8+ T-cell-dependent but NK1.1+ cell-independent. We demonstrated the ability of DCs to produce peroxynitrites and to kill tumor cells; this killing activity involved peroxynitrites. Altogether, these findings make killer DCs the pivotal actors in the beneficial clinical outcome that accompanies antitumor immune responses. We asked whether efficacy can be improved by combining DC-vaccination with the FOLFIRINOX regimen. Combined treatment significantly increased the lifespan of KIC mice with PAC. Prolonged treatment with FOLFIRINOX clearly augmented this beneficial effect. Combining iDC-vaccination with FOLFIRINOX may therefore represent a promising therapeutic option for patients with PAC.- Published
- 2018
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18. Cadherin-1 and cadherin-3 cooperation determines the aggressiveness of pancreatic ductal adenocarcinoma.
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Siret C, Dobric A, Martirosyan A, Terciolo C, Germain S, Bonier R, Dirami T, Dusetti N, Tomasini R, Rubis M, Garcia S, Iovanna J, Lombardo D, Rigot V, and André F
- Subjects
- Animals, Antigens, CD genetics, Cadherins genetics, Carcinoma, Pancreatic Ductal genetics, Cell Line, Tumor, Cell Membrane metabolism, Cell Movement, Cell Proliferation, Collagen Type I metabolism, Disease Progression, Humans, Mice, Neoplasm Transplantation, Pancreatic Neoplasms genetics, Antigens, CD metabolism, Cadherins metabolism, Carcinoma, Pancreatic Ductal metabolism, Pancreatic Neoplasms metabolism
- Abstract
Background: Pancreatic ductal adenocarcinoma (PDAC) is characterised by an extensive tissue invasion and an early formation of metastasis. Alterations in the expression of cadherins have been reported in PDAC. Yet, how these changes contribute to tumour progression is poorly understood. Here, we investigated the relationship between cadherins expression and PDAC development., Methods: Cadherins expression was assessed by immunostaining in both human and murine tissue specimens. We have generated pancreatic cancer cell lines expressing both cadherin-1 and cadherin-3 or only one of these cadherins. Functional implications of such genetic alterations were analysed both in vitro and in vivo., Results: Cadherin-3 is detected early at the plasma membrane during progression of pancreatic intraepithelial neoplasia 1 (PanIN-1) to PDAC. Despite tumoural cells turn on cadherin-3, a significant amount of cadherin-1 remains expressed at the cell surface during tumourigenesis. We found that cadherin-3 regulates tumour growth, while cadherin-1 drives type I collagen organisation in the tumour. In vitro assays showed that cadherins differentially participate to PDAC aggressiveness. Cadherin-3 regulates cell migration, whereas cadherin-1 takes part in the invadopodia activity., Conclusions: Our results show differential, but complementary, roles for cadherins during PDAC carcinogenesis and illustrate how their expression conditions the PDAC aggressiveness.
- Published
- 2018
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19. Saccharomyces boulardii CNCM I-745 Restores intestinal Barrier Integrity by Regulation of E-cadherin Recycling.
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Terciolo C, Dobric A, Ouaissi M, Siret C, Breuzard G, Silvy F, Marchiori B, Germain S, Bonier R, Hama A, Owens R, Lombardo D, Rigot V, and André F
- Subjects
- Cell Line, Cell Membrane Permeability, Colitis, Ulcerative metabolism, Crohn Disease metabolism, Flow Cytometry, Fluorescent Antibody Technique, Humans, Microscopy, Video, Oligonucleotide Array Sequence Analysis, Real-Time Polymerase Chain Reaction, Cadherins metabolism, Intestinal Mucosa metabolism, Saccharomyces boulardii
- Abstract
Background and Aims: Alteration in intestinal permeability is the main factor underlying the pathogenesis of many diseases affecting the gut, such as inflammatory bowel disease [IBD]. Characterization of molecules targeting the restoration of intestinal barrier integrity is therefore vital for the development of alternative therapies. The yeast Saccharomyces boulardii CNCM I-745 [Sb], used to prevent and treat antibiotic-associated infectious and functional diarrhea, may have a beneficial effect in the treatment of IBD., Methods: We analyzed the impact of Sb supernatant on tissue integrity and components of adherens junctions using cultured explants of colon from both IBD and healthy patients. To evaluate the pathways by which Sb regulates the expression of E-cadherin at the cell surface, we developed in vitro assays using human colonic cell lines, including cell aggregation, a calcium switch assay, real-time measurement of transepithelial electrical resistance [TEER] and pulse-chase experiments., Results: We showed that Sb supernatant treatment of colonic explants protects the epithelial morphology and maintains E-cadherin expression at the cell surface. In vitro experiments revealed that Sb supernatant enhances E-cadherin delivery to the cell surface by re-routing endocytosed E-cadherin back to the plasma membrane. This process, involving Rab11A-dependent recycling endosome, leads to restoration of enterocyte adherens junctions, in addition to the overall restoration and strengthening of intestinal barrier function., Conclusion: These findings open new possibilities of discovering novel options for prevention and therapy of diseases that affect intestinal permeability., (Copyright © 2017 European Crohn's and Colitis Organisation (ECCO). Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com)
- Published
- 2017
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20. Interplay between cadherins and α2β1 integrin differentially regulates melanoma cell invasion.
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Siret C, Terciolo C, Dobric A, Habib MC, Germain S, Bonnier R, Lombardo D, Rigot V, and André F
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- Animals, Cell Adhesion, Cell Line, Tumor, Humans, Melanoma metabolism, Mice, Neoplasm Invasiveness, Neoplasm Metastasis, Neoplasm Transplantation, Skin Neoplasms metabolism, Cadherins metabolism, Integrin alpha2beta1 metabolism, Melanoma pathology, Skin Neoplasms pathology
- Abstract
Background: Malignant transformation of melanocytes frequently coincides with an alteration in the expression of cell-cell adhesion molecules (cadherins) and cell-extracellular matrix proteins (integrins). How these two adhesion systems interplay to impact on cell invasion remains to be described in melanoma., Methods: Cell adhesion networks were localised by immunofluorescence in human primary cutaneous melanoma, metastatic melanoma in the lymph nodes, and melanoma cell lines. The role of these cell adhesion networks was assessed both in vivo, by analysing their impact on tumour growth in mice, and in vitro, with the use of functional tests including cell aggregation and cell migration., Results: We found that α2β1 integrin associates with both E-cadherin and N-cadherin to form two adhesive networks, distinguishable by the interaction-or not-of α2β1 integrin with type I collagen. N-cadherin/α2β1 integrin and E-cadherin/α2β1 integrin networks differently participated towards tumour growth in mice. The N-cadherin/α2β1 integrin network showed specific involvement in melanoma cell invasion and migration towards type I collagen. On the other hand, the E-cadherin/α2β1 network regulated cell-cell adhesion., Conclusions: This suggests that different signalling environments can be generated, depending on the type and/or local concentration of cadherin present in the adhesion complex, which potentially leads to differential cell responses. Further clarification of how these adhesive networks are regulated is fundamental to understanding important physiological and pathological processes such as morphogenesis, wound healing, tumour invasion and metastasis.
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- 2015
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21. Saccharomyces boulardii improves intestinal epithelial cell restitution by inhibiting αvβ5 integrin activation state.
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Canonici A, Pellegrino E, Siret C, Terciolo C, Czerucka D, Bastonero S, Marvaldi J, Lombardo D, Rigot V, and André F
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- Animals, Cell Adhesion, Cell Line, Tumor, Cell Movement, Enterocytes cytology, Feeding Behavior, Female, Humans, Mice, Mice, Inbred C57BL, Molecular Weight, Protein Binding, Protein Transport, Signal Transduction, Vitronectin metabolism, Enterocytes metabolism, Enterocytes microbiology, Receptors, Vitronectin metabolism, Saccharomyces physiology
- Abstract
Intestinal epithelial cell damage is frequently seen in the mucosal lesions of infectious or inflammatory bowel diseases such as ulcerative colitis or Crohn's disease. Complete remission of these diseases requires both the disappearance of inflammation and the repair of damaged epithelium. Saccharomyces boulardii (Sb, Biocodex) is a non-pathogenic yeast widely used as a preventive and therapeutic probiotic for the prevention and treatment of diarrhea and other gastrointestinal disorders. We recently showed that it enhances the repair of intestinal epithelium through activation of α2β1 integrin collagen receptors. In the present study, we demonstrated that α2β1 integrin is not the sole cell-extracellular matrix receptor involved during Sb-mediated intestinal restitution. Indeed, by using cell adhesion assays, we showed that Sb supernatant contains heat sensitive molecule(s), with a molecular weight higher than 9 kDa, which decreased αvβ5 integrin-mediated adhesion to vitronectin by competing with the integrin. Moreover, Sb-mediated changes in cell adhesion to vitronectin resulted in a reduction of the αvβ5signaling pathway. We used a monolayer wounding assay that mimics in vivo cell restitution to demonstrate that down-modulation of the αvβ5 integrin-vitronectin interaction is related to Sb-induced cell migration. We therefore postulated that Sb supernatant contains motogenic factors that enhance cell restitution through multiple pathways, including the dynamic fine regulation of αvβ5 integrin binding activity. This could be of major importance in diseases characterized by severe mucosal injury, such as inflammatory and infectious bowel diseases.
- Published
- 2012
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22. αv integrin processing interferes with the cross-talk between αvβ5/β6 and α2β1 integrins.
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Defilles C, Montero MP, Lissitzky JC, Rome S, Siret C, Luis J, André F, and Rigot V
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- Cell Adhesion, Cell Movement, Collagen Type I metabolism, Focal Adhesion Protein-Tyrosine Kinases metabolism, Humans, Mitogen-Activated Protein Kinases metabolism, Phosphatidylinositol 3-Kinase metabolism, Phosphorylation, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction, alpha 1-Antitrypsin biosynthesis, Antigens, Neoplasm metabolism, Integrin alpha2beta1 metabolism, Integrin alphaV metabolism, Integrins metabolism, Receptors, Vitronectin metabolism
- Abstract
Background Information: Previous studies have reported that cross-talk between integrins may be an important regulator of integrin-ligand binding and subsequent signalling events that control a variety of cell functions in many tissues. We previously demonstrated that αvβ5/β6 integrin represses α2β1-dependent cell migration. The αv subunits undergo an endoproteolytic cleavage by protein convertases, whose role in tumoral invasion has remained controversial., Results: Inhibition of convertases by the convertase inhibitor α1-PDX (α1-antitrypsin Portland variant), leading to the cell-surface expression of an uncleaved form of the αv integrin, stimulated cell migration toward type I collagen. Under convertase inhibition, α2β1 engagement led to enhanced phosphorylation of both FAK (focal adhesion kinase) and MAPK (mitogen-activated protein kinase). This outside-in signalling stimulation was associated with increased levels of activated β1 integrin located in larger than usual focal-adhesion structures and a cell migration that was independent of the PI3K (phosphoinositide 3-kinase)/Akt (also called protein kinase B) pathway., Conclusions: The increase in cell migration observed upon convertases inhibition appears to be due to the up-regulation of β1 integrins and to their location in larger focal-adhesion structures. The endoproteolytic cleavage of αv subunits is necessary for αvβ5/β6 integrin to control α2β1 function and could thus play an essential role in colon cancer cell migration.
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- 2011
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23. Saccharomyces boulardii improves intestinal cell restitution through activation of the α2β1 integrin collagen receptor.
- Author
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Canonici A, Siret C, Pellegrino E, Pontier-Bres R, Pouyet L, Montero MP, Colin C, Czerucka D, Rigot V, and André F
- Subjects
- Animals, Caco-2 Cells, Cell Adhesion, Cell Movement drug effects, Enterocytes cytology, Enterocytes drug effects, Enterocytes metabolism, Female, HT29 Cells, Humans, Immunohistochemistry, Mice, Integrin alpha2beta1 metabolism, Probiotics pharmacology, Probiotics therapeutic use, Receptors, Collagen metabolism, Saccharomyces
- Abstract
Intestinal epithelial cell damage is frequently seen in the mucosal lesions of inflammatory bowel diseases such as ulcerative colitis or Crohn's disease. Complete remission of these diseases requires both the cessation of inflammation and the migration of enterocytes to repair the damaged epithelium. Lyophilized Saccharomyces boulardii (Sb, Biocodex) is a nonpathogenic yeast widely used as a therapeutic agent for the treatment and prevention of diarrhea and other gastrointestinal disorders. In this study, we determined whether Sb could accelerate enterocyte migration. Cell migration was determined in Sb force-fed C57BL6J mice and in an in vitro wound model. The impact on α2β1 integrin activity was assessed using adhesion assays and the analysis of α2β1 mediated signaling pathways both in vitro and in vivo. We demonstrated that Sb secretes compounds that enhance the migration of enterocytes independently of cell proliferation. This enhanced migration was associated with the ability of Sb to favor cell-extracellular matrix interaction. Indeed, the yeast activates α2β1 integrin collagen receptors. This leads to an increase in tyrosine phosphorylation of cytoplasmic molecules, including focal adhesion kinase and paxillin, involved in the integrin signaling pathway. These changes are associated with the reorganization of focal adhesion structures. In conclusion Sb secretes motogenic factors that enhance cell restitution through the dynamic regulation of α2β1 integrin activity. This could be of major importance in the development of novel therapies targeting diseases characterized by severe mucosal injury, such as inflammatory and infectious bowel diseases.
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- 2011
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24. Expression of integrin alpha6beta1 enhances tumorigenesis in glioma cells.
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Delamarre E, Taboubi S, Mathieu S, Bérenguer C, Rigot V, Lissitzky JC, Figarella-Branger D, Ouafik L, and Luis J
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- Animals, Apoptosis, Brain Neoplasms metabolism, Cell Line, Tumor, Cell Movement, Cell Proliferation, Female, Glioblastoma metabolism, Humans, Male, Mice, Mice, Inbred BALB C, Neoplasm Invasiveness, Brain Neoplasms pathology, Glioblastoma pathology, Integrin alpha6beta1 biosynthesis
- Abstract
The integrin alpha6beta1 and its main ligand laminin-111 are overexpressed in glioblastoma, as compared with normal brain tissue, suggesting they may be involved in glioblastoma malignancy. To address this question, we stably expressed the alpha6 integrin subunit in the U87 cell line via retroviral-mediated gene transfer. We show that cell surface expression of the alpha6beta1 integrin led to dramatic changes in tumor U87 cell behavior, both in vitro and in vivo. Nude mice receiving either subcutaneous or intracerebral inoculation of alpha6beta1-expressing cells developed substantially more voluminous tumors than mice injected with control cells. The difference in tumor growth was associated with a marked increase in vascularization in response to alpha6beta1 integrin expression and may also be related to changes in the balance between cell proliferation and survival. Indeed, expression of alpha6beta1 enhanced proliferation and decreased apoptosis of U87 cells both in the tumor and in vitro. Additionally, we demonstrate that alpha6beta1 is implicated in glioblastoma cell migration and invasion and that laminin-111 might mediate dissemination of alpha6beta1-positive cells in vivo. Our results highlight for the first time the considerable role of the integrin alpha6beta1 in glioma progression.
- Published
- 2009
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25. alphavbeta5/beta6 integrin suppression leads to a stimulation of alpha2beta1 dependent cell migration resistant to PI3K/Akt inhibition.
- Author
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Defilles C, Lissitzky JC, Montero MP, André F, Prévot C, Delamarre E, Marrakchi N, Luis J, and Rigot V
- Subjects
- Antigens, Neoplasm genetics, Antigens, Neoplasm physiology, Cell Line, Tumor, Cell Movement drug effects, Collagen Type I metabolism, Focal Adhesions, Humans, Integrin alpha2beta1 antagonists & inhibitors, Integrin alpha2beta1 genetics, Integrins genetics, Integrins physiology, Oncogene Protein v-akt genetics, Oncogene Protein v-akt physiology, Phosphatidylinositol 3-Kinases physiology, Protein Kinase Inhibitors pharmacology, RNA, Small Interfering genetics, Receptors, Vitronectin genetics, Receptors, Vitronectin physiology, Signal Transduction, Cell Movement physiology, Integrin alpha2beta1 physiology, Integrins antagonists & inhibitors, Oncogene Protein v-akt antagonists & inhibitors, Phosphoinositide-3 Kinase Inhibitors, Receptors, Vitronectin antagonists & inhibitors
- Abstract
Crosstalk between integrins is involved in the regulation of various cell functions including cell migration. Here we identify the interplay between the integrins alphavbeta5/beta6 and alpha2beta1 during cell migration toward type I collagen. Human colon cancer cell lines HT29-D4 and SW480 were used as cell models. To improve our understanding of the consequences of alphavbeta5/beta6 function on alpha2beta1, we decreased the expression of alphav integrins by either siRNA or lysosomal targeting strategies or inhibited their function using, as antagonists, blocking antibodies or disintegrins. In all cases, we observed a greatly enhanced alpha2beta1 integrin-dependent cell migration associated with focal adhesion rearrangements and increased outside-in signaling as demonstrated by elevated phosphorylation of focal adhesion kinase and MAPKinase (ERK1 and ERK2). The alphavbeta5/beta6-dependent limitation of alpha2beta1 function could be overridden by TS2/16, an activating anti-beta1 antibody. Interestingly, compared to control cells, the pharmacological inhibition of PI3Kinase or the siRNA-mediated knockdown of AKT had little effect on the high alpha2beta1-mediated cell migration observed in the absence of alphav integrins or following activation of alpha2beta1 integrins by the TS2/16. These results suggest that integrins alphavbeta5/beta6 repress alpha2beta1 possibly by interfering with their activation process and thereby modify the cell signaling regulation of alpha2beta1-mediated migration.
- Published
- 2009
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26. Insulin-like growth factor-I receptor, E-cadherin and alpha v integrin form a dynamic complex under the control of alpha-catenin.
- Author
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Canonici A, Steelant W, Rigot V, Khomitch-Baud A, Boutaghou-Cherid H, Bruyneel E, Van Roy F, Garrouste F, Pommier G, and André F
- Subjects
- Cell Adhesion, Cell Movement, Flow Cytometry, Fluorescent Antibody Technique, HT29 Cells metabolism, Humans, Immunoprecipitation, Insulin Receptor Substrate Proteins, Phosphoproteins antagonists & inhibitors, Phosphoproteins genetics, Phosphoproteins metabolism, RNA, Small Interfering pharmacology, Cadherins metabolism, Integrin alphaV metabolism, Receptor, IGF Type 1 metabolism, alpha Catenin pharmacology
- Abstract
Dynamic crosstalk between cell adhesion molecules, extracellular matrix and soluble informative factors is essential for cancer cell migration and invasion. Here, we investigated the mechanisms by which the E-cadherin/catenin complex and alpha v integrin can modulate insulin-like growth factor-I (IGF-I)-induced cell migration. Human colon mucosa, human colon cancer cell lines, HT29-D4 and HCT-8 derivatives that differ in their expression of alpha-catenin, were used as models. Interactions between E-cadherin, alpha v integrin and IGF-I receptor (IGF-IR) were analyzed by coimmunoprecipitation and immunolocalization experiments. The impact of these interactions on cell mobility was determined by haptotaxis assays. We report that alpha v integrin, E-cadherin and IGF-IR form a ternary complex in both cultured cancer cells and human normal colonic mucosa. alpha-Catenin regulates the scaffolding of this complex. IGF-IR ligation by IGF-I induces the disruption of the complex and the relocalization of alpha v integrin from cell-cell contacts to focal contact sites. This perturbation is correlated with the observed increase in cell migration. These results suggest that regulation of the alpha v integrin/E-cadherin/IGF-IR scaffolding is essential for the modulation of cell mobility. Its alteration could be of major importance to sustain alterations in cell adhesion that occur during cancer cell invasion and metastasis., ((c) 2007 Wiley-Liss, Inc.)
- Published
- 2008
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27. Inhibition of proprotein convertases enhances cell migration and metastases development of human colon carcinoma cells in a rat model.
- Author
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Nejjari M, Berthet V, Rigot V, Laforest S, Jacquier MF, Seidah NG, Remy L, Bruyneel E, Scoazec JY, Marvaldi J, and Luis J
- Subjects
- Adenocarcinoma pathology, Adenocarcinoma secondary, Animals, Animals, Genetically Modified, Animals, Newborn, Cell Line, Tumor, Collagen, Humans, Neoplasm Invasiveness, Neoplasm Metastasis prevention & control, Rats, Recombinant Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Transplantation, Heterologous, alpha 1-Antitrypsin metabolism, Cell Movement physiology, Colonic Neoplasms pathology, Neoplasm Metastasis pathology, Proprotein Convertases antagonists & inhibitors, alpha 1-Antitrypsin genetics
- Abstract
Although proprotein convertases are involved in tumor development, nothing is known about their role in metastatic dissemination. To investigate the involvement of convertase inhibition, we used human colon carcinoma cells overexpressing alpha1-antitrypsin Portland (alpha1-PDX, PDX39P cells), a potent convertase inhibitor. We previously reported that these cells bear uncleaved integrin alpha subunits and display an altered attachment to vitronectin that is correlated with defects in the intracellular signaling pathways activated by alphavbeta5 integrin ligation. In this study, we demonstrate that the inhibition of proprotein convertase activity either by overexpression of alpha1-PDX or with the synthetic inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (dec-RVKR-cmk) led to a significant increase in cell migration supported by the alphavbeta5 integrin. A collagen gel invasion assay showed that PDX39P cells also displayed an invasive ability, contrary to control cells. Moreover, when injected to immunosuppressed newborn rats, PDX39P cells were highly invasive, as they induce 10 times more metastases than mock-transfected cells. In addition, the aggressiveness of PDX39P cells can be greatly reduced by a function-blocking monoclonal antibody (mAb) against the alphav subunit. It thus seems that inhibition of proprotein convertases enhances the in vivo invasiveness of colon tumor cells likely due to an increase in cell migration mediated by alphav integrins.
- Published
- 2004
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28. The endoproteolytic processing of alphavbeta5 integrin is involved in cytoskeleton remodelling and cell migration.
- Author
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Berthet V, Rigot V, Nejjari M, Marvaldi J, and Luis J
- Subjects
- Adenocarcinoma, Cell Adhesion drug effects, Cell Adhesion physiology, Cell Line, Cell Movement drug effects, Humans, Laminin pharmacology, Protein Processing, Post-Translational, Signal Transduction drug effects, Signal Transduction physiology, Tetradecanoylphorbol Acetate pharmacology, Transfection, Tumor Cells, Cultured, Vitronectin pharmacology, Cell Movement physiology, Cytoskeleton physiology, Integrins chemistry, Integrins metabolism, Receptors, Vitronectin chemistry, Receptors, Vitronectin metabolism
- Abstract
We previously showed that the post-translational cleavage of alphav subunit is essential for integrin-dependent signalling and cell adhesion. Here, we report that blocking alphav subunit cleavage by expression of alpha1-PDX, a convertase inhibitor, modified the capacity of cells to change shape, via a remodelling of the actin cytoskeleton upon cell attachment. These changes are associated with cell scattering and with a dramatic increase in cell migration to vitronectin. The alphav subunit cleavage is thus essential for integrin function and has a considerable impact on integrin-dependent events, especially those leading to cell migration.
- Published
- 2004
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29. Distinct sites on ABCA1 control distinct steps required for cellular release of phospholipids.
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Rigot V, Hamon Y, Chambenoit O, Alibert M, Duverger N, and Chimini G
- Subjects
- ATP Binding Cassette Transporter 1, ATP-Binding Cassette Transporters genetics, Binding Sites, HeLa Cells, Humans, Immunoblotting, Mutation, Missense, Structure-Activity Relationship, Tangier Disease genetics, ATP-Binding Cassette Transporters metabolism, Cell Membrane metabolism, Phospholipids metabolism
- Abstract
The loss of ABCA1 function leads to Tangier dyslipidemia in humans and to a Tangier-like phenotype in mice, by impairing the transformation of nascent apolipoproteins into mature HDL particles. Mechanistically this ensues from the inability of cells to release membrane lipids and cholesterol. Whereas the ability of ABCA1 to promote phospholipid effluxes, surface binding of apolipoproteins and outward flip of membrane lipids has been documented, the relationship between this series of ABCA1-dependent events is still elusive. Here we provide evidence that i) lipid effluxes require both flip of membrane lipids and binding of apolipoproteins to the cell surface, ii) apolipoprotein A-I binding depends on structural determinants on ABCA1, and iii) phospholipid effluxes can be modulated by engineered mutations on the structural determinants identified on ABCA1.
- Published
- 2002
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30. Protein kinase A site-specific phosphorylation regulates ATP-binding cassette A1 (ABCA1)-mediated phospholipid efflux.
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See RH, Caday-Malcolm RA, Singaraja RR, Zhou S, Silverston A, Huber MT, Moran J, James ER, Janoo R, Savill JM, Rigot V, Zhang LH, Wang M, Chimini G, Wellington CL, Tafuri SR, and Hayden MR
- Subjects
- ATP Binding Cassette Transporter 1, ATP-Binding Cassette Transporters chemistry, Amino Acid Sequence, Animals, Apolipoprotein A-I metabolism, Cells, Cultured, Humans, Mice, Molecular Sequence Data, Phosphorylation, Serine, Structure-Activity Relationship, ATP-Binding Cassette Transporters physiology, Cyclic AMP-Dependent Protein Kinases metabolism, Phospholipids metabolism
- Abstract
ATP-binding cassette A1 (ABCA1) is a key mediator of cholesterol and phospholipid efflux to apolipoprotein particles. We show that ABCA1 is a constitutively phosphorylated protein in both RAW macrophages and in a human embryonic kidney cell line expressing ABCA1. Furthermore, we demonstrate that phosphorylation of ABCA1 is mediated by protein kinase A (PKA) or a PKA-like kinase in vivo. Through site-directed mutagenesis studies of consensus PKA phosphorylation sites and in vitro PKA kinase assays, we show that Ser-1042 and Ser-2054, located in the nucleotide binding domains of ABCA1, are major phosphorylation sites for PKA. ApoA-I-dependent phospholipid efflux was decreased significantly by mutation of Ser-2054 alone and Ser-1042/Ser-2054 but was not significantly impaired with Ser-1042 alone. The mechanism by which ABCA1 phosphorylation affected ApoA-I-dependent phospholipid efflux did not involve either alterations in ApoA-I binding or changes in ABCA1 protein stability. These studies demonstrate a novel serine (Ser-2054) on the ABCA1 protein crucial for PKA phosphorylation and for regulation of ABCA1 transporter activity.
- Published
- 2002
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31. Identification and functional analysis of a naturally occurring E89K mutation in the ABCA1 gene of the WHAM chicken.
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Attie AD, Hamon Y, Brooks-Wilson AR, Gray-Keller MP, MacDonald ML, Rigot V, Tebon A, Zhang LH, Mulligan JD, Singaraja RR, Bitgood JJ, Cook ME, Kastelein JJ, Chimini G, and Hayden MR
- Subjects
- ATP Binding Cassette Transporter 1, ATP-Binding Cassette Transporters metabolism, Animals, Base Sequence, Carotenoids blood, Cell Membrane metabolism, Chickens blood, Cholesterol blood, Chromosome Mapping, Disease Models, Animal, Endoplasmic Reticulum metabolism, HeLa Cells metabolism, HeLa Cells ultrastructure, Humans, Lipoproteins blood, Lipoproteins genetics, Mice, Microscopy, Confocal, Phenotype, Phospholipids blood, Phospholipids genetics, Protein Transport genetics, Sequence Homology, Nucleic Acid, Tangier Disease genetics, ATP-Binding Cassette Transporters genetics, Chickens genetics, Mutation, Missense genetics
- Abstract
The Wisconsin hypoalpha mutant (WHAM) chicken has a >90% reduction in plasma HDL due to hypercatabolism by the kidney of lipid-poor apoA-I. The WHAM chickens have a recessive white skin phenotype caused by a single-gene mutation that maps to the chicken Z-chromosome. This corresponds to human 9q31.1, a chromosomal segment that contains the ATP-binding cassette protein-1 (ABCA1) gene, which is mutated in Tangier Disease and familial hypoalphalipoproteinemia. Complete sequencing of the WHAM ABCA1 cDNA identified a missense mutation near the N-terminus of the protein (E89K). The substitution of this evolutionary conserved glutamate residue for lysine in the mouse ABCA1 transporter leads to complete loss of function, resulting principally from defective intracellular trafficking and very little ABCA1 reaching the plasma membrane. The WHAM chicken is a naturally occurring animal model for Tangier Disease.
- Published
- 2002
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32. In vivo perimenstrual activation of progelatinase B (proMMP-9) in the human endometrium and its dependence on stromelysin 1 (MMP-3) ex vivo.
- Author
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Rigot V, Marbaix E, Lemoine P, Courtoy PJ, and Eeckhout Y
- Subjects
- Animals, Antibodies, Monoclonal metabolism, Blotting, Western, Culture Techniques, Dose-Response Relationship, Drug, Endometrium enzymology, Endometrium metabolism, Enzyme Activation, Enzyme Inhibitors pharmacology, Enzyme Precursors antagonists & inhibitors, Enzyme Precursors biosynthesis, Estradiol metabolism, Female, Gelatinases antagonists & inhibitors, Gelatinases biosynthesis, Humans, Matrix Metalloproteinase 3 biosynthesis, Matrix Metalloproteinase Inhibitors, Metalloendopeptidases antagonists & inhibitors, Metalloendopeptidases biosynthesis, Mice, Progesterone metabolism, Protein Binding, Time Factors, Enzyme Precursors metabolism, Gelatinases metabolism, Matrix Metalloproteinase 3 chemistry, Menstrual Cycle, Metalloendopeptidases metabolism
- Abstract
Most matrix metalloproteinases (MMPs) are secreted as inactive proenzymes. Their expression is well documented in several human tissues, but their activators in vivo are still unknown. To address this question, the activation of progelatinase B (proMMP-9) in the human endometrium was selected as a model system. ProMMP-9 was detected by gelatin zymography in homogenates of fresh endometrial tissue sampled during all phases of the menstrual cycle, whereas its active form was observed only during the late secretory and menstrual phases. Furthermore, proMMP-9 was expressed and activated in endometrial explants sampled outside the perimenstrual phase and cultured in the absence of both progesterone and oestradiol, mimicking the menstrual condition in vivo. Analysis of such tissue cultures by gelatin zymography and Western blotting showed that activation of proMMP-9 depended on a secreted factor and was selectively inhibited by either a synthetic inhibitor of stromelysin 1 (MMP-3) or a monoclonal antibody that specifically blocks MMP-3, thus providing strong evidence for the activation of proMMP-9 in vivo by MMP-3. The activation of proMMP-3 was itself inhibited by a broad-range MMP inhibitor in most cultures, but seemed to involve multiple pathways, implying both serine proteinases and metalloproteinases, which could operate in parallel or sequentially.
- Published
- 2001
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33. Involvement of fibronectin type II repeats in the efficient inhibition of gelatinases A and B by long-chain unsaturated fatty acids.
- Author
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Berton A, Rigot V, Huet E, Decarme M, Eeckhout Y, Patthy L, Godeau G, Hornebeck W, Bellon G, and Emonard H
- Subjects
- Collagen metabolism, Elastin metabolism, Fatty Acids, Unsaturated chemistry, Fibronectins chemistry, Humans, Hydrolysis, Kinetics, Recombinant Proteins chemistry, Recombinant Proteins metabolism, Skin enzymology, Skin metabolism, Fatty Acids, Unsaturated pharmacology, Fibronectins metabolism, Matrix Metalloproteinase Inhibitors, Repetitive Sequences, Amino Acid
- Abstract
The matrix metalloproteinases gelatinase A (MMP-2) and gelatinase B (MMP-9) are implicated in the physiological and pathological breakdown of several extracellular matrix proteins. In the present study, we show that long-chain fatty acids (e.g. oleic acid, elaidic acid, and cis- and trans-parinaric acids) inhibit gelatinase A as well as gelatinase B with K(i) values in the micromolar range but had only weak inhibitory effect on collagenase-1 (MMP-1), as assessed using synthetic or natural substrates. The inhibition of gelatinases depended on fatty acid chain length (with C18 > C16, C14, and C10), and the presence of unsaturations increased their inhibitory capacity on both types of gelatinase. Ex vivo experiments on human skin tissue sections have shown that micromolar concentrations of a long-chain unsaturated fatty acid (elaidic acid) protect collagen and elastin fibers against degradation by gelatinases A and B, respectively. In order to understand why gelatinases are more susceptible than collagenase-1 to inhibition by long-chain fatty acids, the possible role of the fibronectin-like domain (a domain unique to gelatinases) in binding inhibitory fatty acids was investigated. Affinity and kinetic studies with a recombinant fibronectin-like domain of gelatinase A and with a recombinant mutant of gelatinase A from which this domain had been deleted pointed to an interaction of long-chain fatty acids with the fibronectin-like domain of the protease. Surface plasmon resonance studies on the interaction of long-chain fatty acids with the three individual type II modules of the fibronectin-like domain of gelatinase A revealed that the first type II module is primarily responsible for binding these compounds.
- Published
- 2001
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34. Temporal and spatial association of matrix metalloproteinases with focal endometrial breakdown and bleeding upon progestin-only contraception.
- Author
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Galant C, Vekemans M, Lemoine P, Kokorine I, Twagirayezu P, Henriet P, Picquet C, Rigot V, Eeckhout Y, Courtoy PJ, and Marbaix E
- Subjects
- Adult, Blotting, Western, Collagenases metabolism, Endometrium cytology, Female, Humans, Immunohistochemistry, Interleukin-1 metabolism, Matrix Metalloproteinase 2 metabolism, Matrix Metalloproteinase 9 metabolism, RNA, Messenger biosynthesis, Stromal Cells metabolism, beta-N-Acetylhexosaminidases metabolism, Contraceptives, Oral, Hormonal adverse effects, Endometrium metabolism, Levonorgestrel adverse effects, Matrix Metalloproteinases metabolism, Menstruation physiology, Progestins adverse effects
- Abstract
The pathogenesis of irregular endometrial bleeding, the main reason for stopping contraception with progestins only, is unknown. Based on the recent reappraisal of the mechanisms of menstrual bleeding, we hypothesized that matrix metalloproteinases initiate this disorder. Volunteers upon Norplant treatment provided endometrial biopsies at the start of a bleeding episode and during nonbleeding intervals. Focal stromal breakdown, collagen fiber lysis, and collagenase-1 messenger ribonucleic acid were evidenced in most bleeding endometria, but never in the nonbleeding ones. In the breaking down areas, immunolabeling for gelatinase A was strongly increased, and that of progesterone and estrogen receptors was decreased. Explants from bleeding endometria produced high collagenase and gelatinase activities, whereas release from nonbleeding endometria was negligible. Bleeding endometria released more latent and active forms of collagenase-1 and active gelatinases A and B, but less tissue inhibitor of metalloproteinases-1, than nonbleeding endometria. Collagenase-1 release closely correlated with that of interleukin-1alpha. In contrast, N:-acetyl-beta-hexosaminidase and tissue inhibitor of metalloproteinases-2 were similarly released in both groups. Thus, endometrial bleeding occurs together with focal stromal breakdown, collagen lysis, expression and activation of several matrix metalloproteinases, and decreased production of tissue inhibitor of metalloproteinases-1. These results may lead to new pharmacological treatment of this common medical problem.
- Published
- 2000
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35. Role of endoproteolytic processing in the adhesive and signaling functions of alphavbeta5 integrin.
- Author
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Berthet V, Rigot V, Champion S, Secchi J, Fouchier F, Marvaldi J, and Luis J
- Subjects
- Animals, Cell Adhesion, Humans, Mice, Mitogen-Activated Protein Kinase 1 physiology, Mitogen-Activated Protein Kinase 3, Mitogen-Activated Protein Kinases physiology, Protein Processing, Post-Translational, Protein Subunits, Rats, Tumor Cells, Cultured, Vitronectin metabolism, Integrins physiology, Receptors, Vitronectin, Signal Transduction, Subtilisin physiology
- Abstract
Some integrin alpha subunits undergo a post-translational cleavage in their extracellular domain. However, the role of this cleavage in integrin function is unclear. Enzymes involved in this maturation belong to the subtilisin-like endoprotease family (convertases). To understand the role of the alpha subunit cleavage in integrin function, we have designed stable transfectants (PDX39P cells) expressing alpha(1)-PDX, a convertase inhibitor. Immunoprecipitation of cell surface proteins from PDX39P showed that alpha(3), alpha(6) and alpha(v) integrins lack endoproteolytic cleavage. We have compared adhesion between PDX39P cells and mock-transfected cells on different extracellular matrix proteins. No difference in adhesion could be observed on laminin-1 and type I collagen, while attachment of PDX39P cells to vitronectin (ligand of the alpha(v)beta(5) integrin) was dramatically reduced. The reduced adhesion of PDX39P cells was not due to changes in integrin affinity as determined by solid-phase receptor assay in a cell-free environment. Intracellular signaling pathways activated by alpha(v) integrin ligation were also affected in PDX39P cells. It thus seems that the absence of endoproteolytic cleavage of alpha(v) integrins has important consequences on signal transduction pathways leading to alterations in integrin function such as cell adhesion.
- Published
- 2000
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36. Outside-in regulation of integrin clustering processes by ECM components per se and their involvement in actin cytoskeleton organization in a colon adenocarcinoma cell line.
- Author
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Honoré S, Pichard V, Penel C, Rigot V, Prév t C, Marvaldi J, Briand C, and Rognoni JB
- Subjects
- Caco-2 Cells pathology, Cell Adhesion, Cell Movement, Cytoskeletal Proteins metabolism, Fluorescent Antibody Technique, Indirect, Humans, Microscopy, Confocal, Up-Regulation, Actins metabolism, Caco-2 Cells metabolism, Cytoskeleton metabolism, Extracellular Matrix Proteins metabolism, Integrins metabolism
- Abstract
We investigated in a colon adenocarcinoma cell line, the exclusive role of extracellular matrix (ECM) components in the absence of soluble factors regarding the integrin clustering processes, and their implication in cell adhesion, spreading and organization of the actin cytoskeleton. Caco-2 cells were shown to express at the plasma membrane 11 integrins, some of which (e.g. alpha3beta1, alpha5beta1, alpha6beta1/beta4, alpha8beta1 and alpha(v)beta1/beta5/beta6) were identified for the first time in this cell line. Cell adhesion and spreading processes were governed essentially by lamellipodium, the regulation of which was shown to be induced by two types of integrin clustering processes mediated by ECM proteins alone. During these phenomena, alpha2beta1, alpha(v)beta6 and alpha6beta1 integrins, the Caco-2 cell specific receptors of type IV collagen, fibronectin and laminin, respectively, were clustered in small focal complexes (point contacts), whereas alpha(v)beta5, the vitronectin receptor in this cell line, was aggregated in focal adhesions. The two levels of integrin clustering induced only F-actin cortical web formation organized in thin radial and/or circular filaments. We conclude thus that ECM components per se through their action on integrin clustering are involved in cell adhesion, cortical actin cytoskeleton organization and cell spreading.
- Published
- 2000
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37. Integrins and E-cadherin cooperate with IGF-I to induce migration of epithelial colonic cells.
- Author
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André F, Rigot V, Thimonier J, Montixi C, Parat F, Pommier G, Marvaldi J, and Luis J
- Subjects
- Antibodies, Monoclonal pharmacology, Cadherins biosynthesis, Cadherins metabolism, Cell Membrane metabolism, Colon drug effects, Colon physiology, Cytoskeletal Proteins metabolism, Cytoskeleton metabolism, Dose-Response Relationship, Drug, Fluorescent Antibody Technique, HT29 Cells, Humans, Integrins biosynthesis, Integrins immunology, Microscopy, Video, Phosphorylation, Tyrosine metabolism, beta Catenin, Cadherins physiology, Cell Movement drug effects, Colon cytology, Insulin-Like Growth Factor I pharmacology, Integrins physiology, Peptide Fragments pharmacology, Trans-Activators
- Abstract
Although the detailed mechanisms of cell migration remain largely unknown, it is now clear that growth factors and cell adhesion molecules are crucial for this process. We have shown that type I insulin-like growth factor (IGF-I) promotes migration of human colonic tumour cells. Since morphological analysis suggested an involvement of adhesion molecules, we have now examined the role of integrins (cell-matrix adhesion molecules) and E-cadherin/catenins complex (cell-cell adhesion molecules) in the IGF-I-induced migration. Using a monolayer wounding assay, we have determined that, except for alpha2beta1, all of the integrins expressed in HT29-D4 cells are involved in the induced cell migration. Immunofluorescence studies revealed that upon IGF-I stimulation the integrins reorganized at the leading edge of migrating cells. We also demonstrate that E-cadherin is involved in cell migration. A rapid tyrosine phosphorylation of E-cadherin and beta-catenin was detected upon IGF-I stimulation. Tyrosine phosphorylation was associated with reduced membranous expression of E-cadherin and promotion of cell motility, suggesting a regulation of the E-cadherin/catenins complex. This effect can be reversed by incubating cells with tyrosine kinase inhibitors. Taken together, our results suggest that IGF-I promotes colonic cell migration through reorganization of integrin receptors and through modulation of E-cadherin/catenins complex function., (Copyright 1999 Wiley-Liss, Inc.)
- Published
- 1999
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38. Biogenesis of alpha6beta4 integrin in a human colonic adenocarcinoma cell line involvement of calnexin.
- Author
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Rigot V, André F, Lehmann M, Lissitzky JC, Marvaldi J, and Luis J
- Subjects
- Adenocarcinoma pathology, Biological Transport, Calnexin, Colonic Neoplasms pathology, Dimerization, Endoplasmic Reticulum metabolism, Golgi Apparatus metabolism, HT29 Cells, Humans, Hydrolysis, Integrin alpha6beta4, Temperature, Adenocarcinoma metabolism, Antigens, Surface biosynthesis, Calcium-Binding Proteins metabolism, Colonic Neoplasms metabolism, Integrins biosynthesis
- Abstract
The heterodimer alpha6beta4 is a major integrin and the main laminin receptor in epithelia. The alpha6 integrin subunit is proteolytically cleaved, probably by furin, and glycosylated during its biosynthesis. In the present work, we have investigated the kinetics of the assembly process of alpha6beta4 heterodimers in the colonic adenocarcinoma cell line HT29-D4. We demonstrate that the association of alpha6 and beta4 precursors occurs within the ER, while the endoproteolytic cleavage of pro-alpha6 occurs later, probably in the trans-Golgi network. When pro-alpha6 was blocked within the ER by treatment with brefeldin A, its maturation processing was completely prevented without any consequence on its association with beta4 subunit. Low temperature (20 degrees C) also blocked pro-alpha6 maturation, like brefeldin A, but in addition impaired the integrin assembly. Calnexin, an ER resident protein chaperone, was found to be associated with both the alpha6 and beta4 subunit precursors. Our data suggest that calnexin might be responsible for the prolonged retention of pro-alpha6 within the ER compartment and for the defect of integrin subunit association observed at low temperature.
- Published
- 1999
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39. Migration properties of the human ovarian adenocarcinoma cell line IGROV1: importance of alpha(v)beta3 integrins and vitronectin.
- Author
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Carreiras F, Rigot V, Cruet S, Andre F, Gauduchon P, and Marvaldi J
- Subjects
- Cell Movement, Female, Humans, Immunohistochemistry, Signal Transduction physiology, Tumor Cells, Cultured, Adenocarcinoma pathology, Extracellular Matrix Proteins physiology, Ovarian Neoplasms pathology, Receptors, Vitronectin physiology, Vitronectin physiology
- Abstract
Cell migration of ovarian tumoral cells is essential for cell dissemination and for invasion of the submesothelial extracellular matrix (ECM). We have conducted a study of the migratory properties of an ovarian adenocarcinoma cell line (IGROV1) by using 2 distinct methods for the evaluation of cell migration. We found that in a short-term transfilter migration assay, IGROV1 cells migrated toward vitronectin, fibronectin, type IV collagen and laminin in an integrin-dependent manner. When migration was evaluated in a wound healing assay, the restitution of the wounded area was stimulated solely by added, exogenous vitronectin and was almost totally dependent on alpha(v)beta3 integrin function. Moreover, we demonstrated that alpha(v)beta3 was localized in focal contacts restricted to the leading edge of migrating cells, whereas vitronectin notably localized with actin stress fibers and cortical actin. On the other hand, several kinase inhibitors were found to impede migration of IGROV1 induced by vitronectin. It thus appears that alpha(v)beta3-vitronectin interactions lead to the activation of multiple signaling pathway including activation of protein kinase C, phosphatidyl-inositol-3-phosphate kinase and protein tyrosine kinase. The "alpha(v)beta3-vitronectin system" is therefore essential to the migration of human ovarian carcinoma cells.
- Published
- 1999
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40. Protein kinases C-gamma and -delta are involved in insulin-like growth factor I-induced migration of colonic epithelial cells.
- Author
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André F, Rigot V, Remacle-Bonnet M, Luis J, Pommier G, and Marvaldi J
- Subjects
- Calcium-Calmodulin-Dependent Protein Kinases antagonists & inhibitors, Calcium-Calmodulin-Dependent Protein Kinases metabolism, Cell Division drug effects, Cell Movement drug effects, Colon drug effects, Epithelial Cells drug effects, HT29 Cells, Humans, Immunohistochemistry, Insulin pharmacology, Insulin physiology, Insulin-Like Growth Factor Binding Protein 6 metabolism, Insulin-Like Growth Factor I pharmacology, Intestinal Mucosa cytology, Intestinal Mucosa drug effects, Isoenzymes antagonists & inhibitors, Peptide Fragments pharmacology, Protein Kinase C antagonists & inhibitors, Protein Kinase C-delta, Cell Movement physiology, Colon cytology, Epithelial Cells physiology, Insulin-Like Growth Factor I physiology, Isoenzymes physiology, Protein Kinase C physiology
- Abstract
Background & Aims: The mechanisms by which epithelial cells migrate during the repair of damaged colonic mucosa are poorly understood. This study investigated the insulin-like growth factor I (IGF-I) signaling pathway leading to HT29-D4 human colonic epithelial cell line migration., Methods: IGF-stimulated cell migration was determined using a wound model in the presence or absence of kinase inhibitors. Activation of protein kinase C (PKC) was determined by immunodetection., Results: IGF-I and insulin induce cell migration without affecting cell proliferation through their cognate receptors. Des(1-3)-IGF-I, a truncated analogue of IGF-I, was more potent than IGF-I, suggesting that IGF-binding proteins secreted in the medium modulated IGF-I-induced cell migration. Inhibition of phosphatidylinositol 3-kinase, PKC, and mitogen-activated protein kinases eliminated cell restitution. Long-term exposure of cells to phorbol myristate acetate caused the depletion of PKC-delta and -gamma and prevented also IGF-I-induced cell motility. IGF-I also induced activation of PKC-delta and -gamma only., Conclusions: IGF-I stimulates colonic restitution through the activation of multiple signaling pathways including activation of phosphatidylinositol 3-kinase, PKC-delta and -gamma, and mitogen-activated protein kinases.
- Published
- 1999
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41. Convergent effects of growth factors, hormones, and fibronectin are necessary for the enterocyte differentiation of a colon adenocarcinoma cell line (HT29-D4).
- Author
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Rognoni JB, Pichard V, Honore S, Rigot V, Lehmann M, Roccabianca M, Carles G, Luis J, Marvaldi J, and Briand C
- Subjects
- Animals, Cattle, Cell Differentiation drug effects, Cell Division drug effects, Culture Media, Serum-Free, Fluorescent Antibody Technique, Indirect, HT29 Cells, Humans, Insulin pharmacology, Intestinal Mucosa pathology, Serum Albumin, Bovine pharmacology, Triiodothyronine pharmacology, Adenocarcinoma pathology, Colonic Neoplasms pathology, Fibronectins pharmacology, Growth Substances pharmacology, Hormones pharmacology, Intestinal Mucosa drug effects
- Abstract
The aim of this work was to show in serum-free medium a convergent effect of physiological factors and extracellular matrix proteins on the differentiation process of enterocytes by taking as a model the HT29-D4 clone that has the feature of differentiating when subcultured in fetal bovine serum glucose-free medium. We show that triiodothyronine (T3) as well as insulin promotes limited cell growth and differentiation, whereas fibronectin or bovine serum albumin (BSA) induces cell growth and a low level of differentiation. However, insulin, T3, fibronectin, and BSA together with epidermal growth factor and transferrin promoted satisfactory growth and enterocyte morphology with epithelial electrophysiological properties in HT29-D4 cells. With these factors adequate protein targeting was achieved since cells apically expressed the carcinoembryonic antigen, and basolaterally transferrin and insulin receptors, beta 1 and alpha v beta 6 integrins, talin, vinculin, and focal adhesion kinase (FAK). Talin, vinculin, FAK, and alpha v beta 6 integrin, the fibronectin receptor, were clustered in focal contacts, which agrees with a possible role of fibronectin in final cell growth, the latter process mediating the final phase of differentiation. This level of differentiation can be maintained for a long time. Thus HT29-D4 cells appear to be a suitable model to study the implication of integrins in the differentiation process of human enterocytes.
- Published
- 1998
- Full Text
- View/download PDF
42. Lack of integrin alpha-chain endoproteolytic cleavage in furin-deficient human colon adenocarcinoma cells LoVo.
- Author
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Lehmann M, Rigot V, Seidah NG, Marvaldi J, and Lissitzky JC
- Subjects
- Adenocarcinoma, Amidohydrolases metabolism, Colonic Neoplasms, Exotoxins toxicity, Furin, Humans, Hydrolysis, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase, Protein Processing, Post-Translational, Recombinant Proteins metabolism, Recombinant Proteins pharmacology, Subtilisins pharmacology, Tumor Cells, Cultured, Pseudomonas aeruginosa Exotoxin A, ADP Ribose Transferases, Bacterial Toxins, Integrins metabolism, Subtilisins metabolism, Virulence Factors
- Abstract
In the present report the biosynthesis of the integrin alpha-chains endowed with constitutive endoproteolytic cleavage was evaluated in LoVo cells where furin, a subtilisin-like convertase involved in post-translational endoproteolytic processing, is not functional. It was found that cell-surface alpha 3, alpha 6 and alpha v subunits were not processed endoproteolytically into heavy and light chains as they were in HT29-D4 cells, a furin-competent cell line. Complete removal of N-linked oligosaccharides and pulse-chase experiments confirmed that the cleavage of the alpha 6 integrin subunit occurring 45 min after translation in HT29 cells did not take place in LoVo cells. Apart from cleavage deficiency, alpha 6 subunit glycosylation, association with beta 4 subunits and targeting to the plasma membrane seemed comparable in LoVo and HT29 cells. The pro-alpha 6 and the pro-alpha 3 subunits immunopurified from LoVo cells were highly sensitive to endoproteolysis by recombinant furin. Furin cleavage was calcium dependent and resulted in the conversion of the 140 kDa pro-alpha 6 into a 120 kDa heavy chain. These results suggest strongly that furin is involved in the endoproteolytic processing of cleavable integrin alpha subunits.
- Published
- 1996
- Full Text
- View/download PDF
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