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1.
Exp Gerontol ; 126: 110693, 2019 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-31430521

RESUMO

BACKGROUND: Glypicans (GPCs) are heparan sulfate cell membrane proteoglycans containing glycosylphosphatidylinositol (GPI) anchor. They play important role in cell behavior by activating/presenting numerous growth factors and cytokines. OBJECTIVES: The expression of GPCs was investigated in primary culture of skin keratinocytes sampled from healthy donors of different age. MATERIALS AND METHODS: Primary keratinocytes from healthy female donors aged from 20 to 89 years old (n = 30) were either isolated from breast or abdominal skin samples (n = 27) or purchased (n = 3). GPCs expression was examined by qPCR, immunohistochemistry and western blot. Its role in proliferation induced by fibroblast growth factor 2 (FGF2) was also studied. RESULTS: Glypican 1 (GPC1) was the major expressed GPC in human keratinocytes. Its expression was up to two orders of magnitude higher than other GPCs and was significantly decreased with the age of the donors. It was localized at the cell surface and associated with intracellular granules. In skin sections, GPC1 was mainly localized in basal layer of epidermis. Shedding of GPCs decreased the proliferative effect of FGF2, confirming their role of modulator of growth factor effects on keratinocytes. These results established GPC1 as an important player in epidermis biology and skin ageing.


Assuntos
Envelhecimento/metabolismo , Glipicanas/metabolismo , Queratinócitos/metabolismo , Pele/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/genética , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Células Cultivadas , Epiderme/metabolismo , Feminino , Fator 2 de Crescimento de Fibroblastos/farmacologia , Regulação da Expressão Gênica/fisiologia , Glipicanas/genética , Humanos , Queratinócitos/efeitos dos fármacos , Pessoa de Meia-Idade , RNA Mensageiro/genética , Transdução de Sinais/fisiologia , Adulto Jovem
2.
Anal Biochem ; 482: 16-21, 2015 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-25935259

RESUMO

Basement membrane collagens or derived fragments are measured in biological fluids such as blood and urine of patients and appear to be useful for diagnosis, prognostication, or treatment monitoring as proposed for endostatin, a fragment of collagen XVIII, or tumstatin, a fragment of collagen IV. Tetrastatin, the NC1 alpha 4 collagen IV domain, was previously reported to inhibit tumor growth and angiogenesis. The aim of this study was to develop and validate a method to measure tetrastatin concentrations in human fluids. We developed a competitive enzyme-linked immunosorbent assay (ELISA). It allowed measuring tetrastatin levels in human serum, bronchial aspiration and bronchoalveolar lavage fluids, and lung tissue extracts. The tetrastatin level was significantly higher in tumor tissues than in healthy lung tissues. Tetrastatin competitive ELISA could be useful to quantify tetrastatin in tissues and biological fluids for the diagnosis or prognostication of diseases in which basement membrane metabolism may be altered, especially tumor progression.


Assuntos
Colágeno Tipo IV/análise , Colágeno Tipo IV/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/diagnóstico , Pulmão/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Líquido da Lavagem Broncoalveolar/química , Criança , Feminino , Humanos , Pulmão/química , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estrutura Terciária de Proteína , Adulto Jovem
3.
Anal Biochem ; 437(2): 111-7, 2013 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-23499763

RESUMO

Type XIX collagen is a minor collagen associated with basement membranes in vascular, neuronal, mesenchymal, and epithelial tissues. We demonstrated that the NC1, C-terminal, domain of collagen XIX inhibits the migration capacities of tumor cells and exerts a strong inhibition of tumor growth. Other basement membrane collagens or derived fragments were measured in biological fluids such as blood and urine of patients and appeared to be useful for diagnosis, prognosis, or treatment monitoring. The aim of this study was to develop and validate methods to measure collagen XIX and its fragments in human cell cultures, tissue extracts, and human biological fluids. For that purpose, we developed real-time PCR, Western blot, and competitive enzyme-linked immunosorbent assays. We demonstrated that the methods developed in this paper are specific for collagen XIX. We showed that it is expressed in human cell cultures, tissue extracts, and various biological fluids. These methods may be used in various human tissue extracts and biological fluids such as serum, amniotic fluid, cord blood, and many other fluids. Collagen XIX or its fragments could constitute new biomarkers for human diseases as well as for diagnosis and/or prognosis.


Assuntos
Líquidos Corporais/química , Colágeno/classificação , Colágeno/metabolismo , Células Epiteliais/metabolismo , Fibroblastos/metabolismo , Extratos de Tecidos/química , Linhagem Celular , Colágeno/química , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/química , Fibroblastos/química , Regulação da Expressão Gênica/fisiologia , Humanos , Osteossarcoma/química , Osteossarcoma/metabolismo
4.
J Physiol Pharmacol ; 60 Suppl 4: 15-22, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20083847

RESUMO

BACKGROUND: Lumican is a small leucine-rich proteoglycan (SLRP) of the extracellular matrix (ECM) involved in the control of melanoma growth and invasion. The aim of the present study was to analyse the role of lumican in the regulation of the development of lung metastasis. METHODS: B16F1 melanoma cells stably transfected with lumican expressing plasmid (Lum-B16F1) were injected to syngenic mice. The lung metastasis was compared to mice injected with mock-transfected B16F1 cells (Mock-B16F1). The expression of lumican, cyclin D1, apoptotic markers, vascular endothelium growth factor (VEGF) and Von Willebrand Factor (vWF) within lung metastasis nodules was investigated by immunohistochemistry. In parallel, cells cultured in presence of lumican were assayed for apoptosis and motility. RESULTS: We observed that the number and the size of lung metastasis nodules were significantly decreased in mice injected with Lum-B16F1 cells in comparison to Mock-B16F1 cells. This was associated with an increase of tumour cell apoptosis within metastasis nodules but the cell proliferation rate remained constant in the two mice groups. In contrast, the VEGF immunostaining and the number of blood vessels within the lung metastasis nodules were decreased in the lumican-expressing tumours. In vitro, a significant decrease of apoptotic markers in wild type B16F1 cells incubated with increasing amounts of lumican core protein was observed. In addition, pseudotubes formation on Matrigel(R) and the migratory capacity of endothelial cells was inhibited by lumican. Altogether, our results indicate that lumican decreases lung metastasis development not only by inducing tumour cell apoptosis but also by inhibiting angiogenesis.


Assuntos
Antineoplásicos , Proteoglicanas de Sulfatos de Condroitina/farmacologia , Sulfato de Queratano/farmacologia , Neoplasias Pulmonares/prevenção & controle , Neoplasias Pulmonares/secundário , Melanoma Experimental/tratamento farmacológico , Inibidores da Angiogênese/farmacologia , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Células Endoteliais/metabolismo , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Lumicana , Neoplasias Pulmonares/patologia , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/farmacologia , Fator A de Crescimento do Endotélio Vascular/biossíntese
5.
Clin Exp Dermatol ; 32(4): 405-16, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17490399

RESUMO

BACKGROUND: The family of small leucine-rich proteoglycans (SLRPs), which includes decorin, lumican, biglycan and fibromodulin, constitutes an abundant component of the skin extracellular matrix. We previously demonstrated that human lumican inhibits melanoma growth and progression in a mouse experimental model, by regulating cell migration, proliferation and apoptosis. AIM: The aim of this study was to investigate the expression of lumican and decorin in human malignant melanoma and adjacent peritumoral tissue, to understand better their role in the control of growth and invasion of human melanoma. METHODS: Expression of both proteoglycans was studied by immunohistochemistry using specific antibodies in 34 malignant melanomas, 12 Hutchinson's melanotic freckles and 4 cutaneous metastatic melanomas. RESULTS: We showed that lumican and decorin are located in the dermis and in the peritumoral stroma of malignant melanoma, but are not found in melanoma cells or dense tumour tissue. In the healthy dermis, distant from the tumour, the increasing ratio of lumican to decorin was inversely correlated with the proliferation of the tumour cells (P = 0.035). The comparison of the level of expression of lumican protein in superficial vs. nodular subtypes of malignant melanomas showed a decrease of lumican but not decorin in the peritumoral stroma of nodular subtypes. In the peritumoral stroma, the level of expression of lumican but not decorin decreased significantly (P = 0.016) with increasing Clark levels. In addition, immunocytochemical and reverse transcription PCR analyses of malignant melanoma cell lines (A-375, HT-144) and of MRC-5 and dermal fibroblasts from healthy donors in vitro confirmed that dermal fibroblasts are responsible for lumican and decorin synthesis in skin. CONCLUSIONS. Lumican may regulate vertical progression of human malignant melanoma, but further study is necessary to clarify the antitumour mechanism and the downstream signal transduction pathways involved.


Assuntos
Proteoglicanas de Sulfatos de Condroitina/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Sulfato de Queratano/metabolismo , Melanoma/metabolismo , Proteínas de Neoplasias/metabolismo , Proteoglicanas/metabolismo , Neoplasias Cutâneas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Decorina , Feminino , Humanos , Imuno-Histoquímica , Lumicana , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Estromais/metabolismo
6.
Ann Biol Clin (Paris) ; 63(5): 553-5, 2005.
Artigo em Francês | MEDLINE | ID: mdl-16230296

RESUMO

Preanalytical steps may be the source of many errors. In this paper, we report the case of an exploration of cerebrospinal fluid (CSF) proteins in which the pre-analytical step was defective. Discordance in the results of the CSF protein level measurement, associated with an aberrant electrophoresis led us to suspect a preanalytical interference. After investigating the preanalytical treatment of the sample, we suspected a possible interference by previous formaldehyde treatment, which was confirmed by several tests performed in our laboratory. These data point out the importance of preanalytical steps for the quality of results.


Assuntos
Proteínas do Líquido Cefalorraquidiano/análise , Eletroforese/métodos , Esclerose/líquido cefalorraquidiano , Adulto , Feminino , Formaldeído , Humanos , Indicadores e Reagentes , Valores de Referência , Reprodutibilidade dos Testes , Esclerose/diagnóstico
7.
Ann Biol Clin (Paris) ; 60(5): 565-70, 2002.
Artigo em Francês | MEDLINE | ID: mdl-12368142

RESUMO

The interest of serum protein immunofixation in myeloma and Waldenstr m's macroglobulinemia is widely known. It is not so well defined in other malignant hemopathies. The purpose of this study was to determine immunofixation abnormalities in malignant hemopathies other than multiple myeloma and Waldenstr m's macroglobulinemia. We selected serum immunofixations of 61 patients affected by malignant hemopathies and 53 patients affected by other pathologies susceptible to give immunofixation's alterations. We showed that the frequency of immunofixation abnormalities was more important in patients affected by malignant hemopathies than in patients affected by other pathologies (70.5% vs 35.8%). A high frequency of monoclonal immunoglobulins was found in patients with lymphoma (53.3%) and oligoclonal immunoglobulins in other hemopathies (48.2%). No significant difference of the frequency of the monoclonal immunoglobulin isotypes was found. In summary, this retrospective study demonstrates a high frequency of immunofixation abnormalities in malignant hemopathies other than multiple myeloma and Waldenstr m's macroglobulinemia and different immunofixation characteristics between lymphomas and other hemopathies.


Assuntos
Eletroforese das Proteínas Sanguíneas/métodos , Eletroforese em Gel de Ágar/métodos , Immunoblotting/métodos , Imunoeletroforese/métodos , Leucemia Linfocítica Crônica de Células B/sangue , Leucemia Linfocítica Crônica de Células B/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/sangue , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Linfoma/sangue , Linfoma/diagnóstico , Idoso , Anticorpos Monoclonais/sangue , Eletroforese das Proteínas Sanguíneas/normas , Estudos de Casos e Controles , Eletroforese em Gel de Ágar/normas , Feminino , Humanos , Immunoblotting/normas , Imunoeletroforese/normas , Imunoglobulina G/sangue , Isotipos de Imunoglobulinas/sangue , Imunoglobulina M/sangue , Imunoglobulinas/sangue , Leucemia Linfocítica Crônica de Células B/imunologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/imunologia , Linfoma/imunologia , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/sangue , Mieloma Múltiplo/diagnóstico , Mieloma Múltiplo/imunologia , Bandas Oligoclonais , Macroglobulinemia de Waldenstrom/sangue , Macroglobulinemia de Waldenstrom/diagnóstico , Macroglobulinemia de Waldenstrom/imunologia
8.
Med Sci Sports Exerc ; 33(4): 628-34, 2001 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11283440

RESUMO

PURPOSE: The purpose of this study was to evaluate physical performance (static and dynamic) of U.S. Air Force reservists after an eastbound air travel across seven time zones and to estimate the pharmacological aids slow-release caffeine and melatonin versus placebo in attempt to overcome the decline in performance. METHODS: 27 American volunteers were randomly divided into three groups: caffeine 300 mg, melatonin 5 mg, and placebo (lactose capsules). Two days before the flight and 10 d after, three tests were performed: hand grip strength test (static performance), squat jump test (maximal height), and multiple jump test (power and endurance). All measures were repeated twice a day: morning and afternoon. RESULTS: In placebo conditions, the static performance of the dominant hand decreased significantly during the first three mornings and tended to decrease the fourth morning. Simultaneously, the caffeine group's static performance increased significantly, whereas the melatonin group maintained its levels. No significant differences were observed the afternoons. No statistical differences appeared for the nondominant hand in the mornings or afternoons. Dynamic capacities presented no significant degradation after the travel. In the placebo group, for the squat jump test, performance increased from the fourth day. No real explanation can be given about this result. CONCLUSIONS: We demonstrated that slow-release caffeine and melatonin might be used to compensate for jet-lag troubles and particularly for the static physical performance decrease. The slow-release caffeine seems to be the best treatment, but its effects are only demonstrated on previously damaged performance. These preliminary results need further investigation, but we are the first to report a beneficial effect of slow-release caffeine and melatonin on physical performances after jet-lag.


Assuntos
Antioxidantes/uso terapêutico , Cafeína/uso terapêutico , Estimulantes do Sistema Nervoso Central/uso terapêutico , Síndrome do Jet Lag/prevenção & controle , Melatonina/uso terapêutico , Adulto , Análise de Variância , Ritmo Circadiano/efeitos dos fármacos , Preparações de Ação Retardada/uso terapêutico , Método Duplo-Cego , Teste de Esforço/métodos , Feminino , Força da Mão/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade
9.
Lab Invest ; 80(8): 1337-43, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10950124

RESUMO

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.


Assuntos
Interleucina-4/farmacologia , Interleucina-4/fisiologia , Cicatrização/fisiologia , Animais , Sequência de Bases , Interleucina-4/genética , Camundongos , Oligonucleotídeos Antissenso/farmacologia
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