Biomedical subjects

Agnès Mailloux

Publications and source records attributed to Agnès Mailloux.

4 recordsLinked to original sources

PubMed2005

Proteomics of human umbilical vein endothelial cells applied to etoposide-induced apoptosis.

We have undertaken to continue the proteomic study of human umbilical vein endothelial cells (HUVECs) using the combination of 2-DE, automated trypsin digestion, and PMF analysis after MALDI-TOF MS and peptide sequencing using nano LC-ESI-MS/MS. The overall functional characterization of the 162 identified proteins from primary cultures of HUVECs confirms the metabolic capabilities of endothelium and illustrates various cellular functions more related to cell motility and angiogenesis, protein folding, anti-oxidant defenses, signal transduction, proteasome pathway and resistance to apoptosis. In comparison with controls cells, the differential proteomic analysis of HUVECs treated by the pro-apoptotic topoisomerase inhibitor etoposide further revealed the variation of eight proteins, namely, GRP78, GRP94, valosin-containing protein, proteinase inhibitor 9, cofilin, 37-kDa laminin receptor protein, bovine apolipoprotein, and tropomyosin. These data suggest that etoposide-induced apoptosis of human vascular endothelial cells results from the intricate involvement of multiple apoptosis processes including at least the mitochondrial and the ER stress pathways. The presented 2-D pattern and protein database, as well as the data related to apoptosis of HUVECs, are available at http://www.huvec.com.

Apoptosis↗

PubMed2003

Proteomic study of human umbilical vein endothelial cells in culture.

The endothelium is a single layer of cells lining the inside face of all blood vessels. It constitutes a major metabolic organ which is critically involved in the generation and the regulation of multiple physiological and pathological processes such as coagulation, hemostasis, inflammation, atherosclerosis, angiogenesis and cancerous metastasis dissemination. In order to increase our knowledge about the protein content and the main biological pathways of human vascular endothelial cells, we have undertaken the proteomic analysis of the most explored present endothelial cell model, i.e. primocultures of human umbilical vein endothelial cells (HUVECs). Using low levels of protein loads (~ 30 nug), the association of two-dimensional electrophoresis with matrix-assisted laser desorption/ionization-time of flight mass spectrometry, liquid chromatography-tandem mass spectrometry and database interrogations allowed us to identify 53 proteins of suspected endothelial origin in quiescent HUVECs. Beside cytoskeletal proteins such as actin, tubulin, tropomyosin and vimentin, we identified various proteins more especially implicated in cellular motility and plasticity (e.g. cofilin, F-actin capping protein and prefoldin), in regulation of apoptosis and senescence (protease inhibitor 9, glucose related proteins, heat shock proteins, thioredoxin peroxidase, nucleophosmin) as well as other proteins implicated in coagulation (annexin V, high mobility group protein), antigen presentation (valosin containing protein and ubiquitin carboxyl terminal hydrolase isozyme L1) and enzymatic capabilities (glutathione-S-transferase, protein disulfide isomerases, lactate deshydrogenase). The presented annotated 2-D maps of HUVECs will be soon available on the web at http://www. huvec.com.

Cells, Cultured↗

PubMed2003

Captopril and enalaprilat decrease antioxidant defences in human endothelial cells and are unable to protect against apoptosis.

Angiotensin-converting enzyme (ACE) inhibitors were shown to improve endothelial dysfunction in various human diseases and some of these inhibitors have been proposed as enhancers of antioxidant defences. We measured glutathione peroxidase (GPX), superoxide dismutase (SOD) and malondialdehyde (MDA) in human endothelial cells treated with captopril or enalaprilat, two ACE inhibitors, and we showed that both inhibitors decreased GPX and SOD activities but not MDA, the end-product of lipoperoxidation. Captopril and enalaprilat were also unable to protect against etoposide-induced apoptosis in endothelial cells, indicating that they cannot be considered as protective drugs for the endothelium, in particular in clinical situations involving oxidative stress or apoptosis. Moreover, when used at high concentration captopril, but not enalaprilat, was toxic for endothelial cells with both necrotic and apoptotic effects.

Angiotensin-Converting Enzyme Inhibitors↗

PubMedDate not supplied

Cyclosporin A but not estradiol can protect endothelial cells against etoposide-induced apoptosis.

This study was undertaken to examine the possibilities of endothelial protection toward toxicity of anticancer drugs. To test the hypothesis that estradiol (E2) and cyclosporin A (CsA) can interfere within programmed cell death in human umbilical vein endothelial cells (HUVECs), apoptosis was induced by etoposide with and without E2 or CsA. All the concentrations of E2 tested (from 10(-9) to 10(-5) M) failed to protect HUVECs. For CsA a dual effect was observed: used at 1 or 10 microg/mL in coincubation with etoposide, CsA significantly reduced etoposide-induced apoptosis but complete inhibition was not reached, whereas used at 50 microg/mL CsA did not protect HUVECs anymore and even had deleterious effects. Furthermore, a 24-h pretreatment of HUVECs by CsA at 10 microg/mL significantly protected the cells by preventing both bcl-2 level decrease and caspase-3 activation related to etoposide-induced apoptosis. Protective effects of CsA toward endothelial cells were concentration dependent; in pretreatment at 10 microg/mL, CsA was an effective protector and might contribute in vivo to inhibit obvious toxic effects caused by anticancer drugs.

Antineoplastic Agents, Phytogenic↗