Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Benzo[a]pyrene (B[a]P), the prototype molecule of polycyclic aromatic hydrocarbons, exhibits genotoxic and carcinogenic effects, which has led the International Agency for Research on Cancer to recognize it as a human carcinogen. Besides the well-known apoptotic signals triggered by B[a]P, survival signals have also been suggested to occur, both signals likely involved in cancer promotion. Our previous work showed that B[a]P induced an hyperpolarization of mitochondrial membrane potential (ΔΨm) in rat hepatic epithelial F258 cells. Elevated ΔΨm plays a role in tumor development and progression, and nitric oxide (NO) has been suggested to be responsible for increases in ΔΨm. The present study therefore aimed at evaluating the impact of B[a]P on NO level in F258 cells, and at testing the putative role for NO as a survival signal, notably in link with ΔΨm. Our data demonstrated that B[a]P exposure resulted in an NO production which was dependent upon the activation of the inducible NO synthase. This enzyme activation involved AhR and possibly p53 activation. Preventing NO production not only increased B[a]P-induced cell death but also blocked mitochondrial hyperpolarization. This therefore points to a role for NO as a survival signal upon B[a]P exposure, possibly targeting ΔΨm
ISSN: 0887-2333

hal-01163757
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01163757
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01163757/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01163757/file/Benzo%5Ba%5Dpyrene-induced%20nitric.pdf
DOI : 10.1016/j.tiv.2015.06.010

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
The mycotoxin enniatin B (EnnB) is predominantly produced by species of the Fusarium genera, and often found in grain. The cytotoxic effect of EnnB has been suggested to be related to its ability to form ionophores in cell membranes. The present study examines the effects of EnnB on cell death, differentiation, proliferation and pro-inflammatory responses in the murine monocyte-macrophage cell line RAW 264.7. Exposure to EnnB for 24 h caused an accumulation of cells in the G0/G1-phase with a corresponding decrease in cyclin D1. This cell cycle-arrest was possibly also linked to the reduced cellular ability to capture and internalize receptors as illustrated by the lipid marker ganglioside GM1. EnnB also increased the number of apoptotic, early apoptotic and necrotic cells, as well as cells with elongated spindle-like morphology. The Neutral Red assay indicated that EnnB induced lysosomal damage; supported by transmission electron microscopy (TEM) showing accumulation of lipids inside the lysosomes forming lamellar structures/myelin bodies. Enhanced levels of activated caspase-1 were observed after EnnB exposure and the caspase-1 specific inhibitor ZYVAD-FMK reduced EnnB-induced apoptosis. Moreover, EnnB increased the release of interleukin-1 beta (IL-1β) in cells primed with lipopolysaccharide (LPS), and this response was reduced by both ZYVAD-FMK and the cathepsin B inhibitor CA-074Me. In conclusion, EnnB was found to induce cell cycle arrest, cell death and inflammation. Caspase-1 appeared to be involved in the apoptosis and release of IL-1β and possibly activation of the inflammasome through lysosomal damage and leakage of cathepsin B.
ISSN: 0041-008X

inserm-00871698
http://www.hal.inserm.fr/inserm-00871698
DOI : 10.1016/j.taap.2012.03.014
PUBMED : 22483798

Éditeur(s) : HAL CCSD
Résumé : TNF-Related Apoptosis-Inducing Ligand (TRAIL) is a well-known apoptosis inducer, which activates the extrinsic death pathway. TRAIL is pro-apoptotic on colon cancer cells, while not cytotoxic towards normal healthy cells. However, its clinical use is limited by cell resistance to cell death which occurs in approximately 50% of cancer cells. Short Chain Fatty Acids (SCFA) are also known to specifically induce apoptosis of cancer cells. In accordance, we have shown that food grade dairy propionibacteria induce intrinsic apoptosis of colon cancer cells, via the production and release of SCFA (propionate and acetate) acting on mitochondria. Here, we investigated possible synergistic effect between Propionibacterium freudenreichii and TRAIL. Indeed, we hypothesized that acting on both extrinsic and intrinsic death pathways may exert a synergistic pro-apoptotic effect. Whole transcriptomic analysis demonstrated that propionibacterial supernatant or propionibacterial metabolites (propionate and acetate), in combination with TRAIL, increased pro-apoptotic gene expression (TRAIL-R2/DR5) and decreased anti-apoptotic gene expression (FLIP, XIAP) in HT29 human colon cancer cells. The revealed synergistic pro-apoptotic effect, depending on both death receptors (TRAIL-R1/DR4, TRAIL-R2/DR5) and caspases (caspase-8, -9 and -3) activation, was lethal on cancer cells but not on normal human intestinal epithelial cells (HIEC), and was inhibited by Bcl-2 expression. Finally, milk fermented by P. freudenreichii induced HT29 cells apoptosis and enhanced TRAIL cytotoxic activity, as did P. freudenreichii DMEM culture supernatants or its SCFA metabolites. These results open new perspectives for food grade P. freudenreichii-containing products in order to potentiate TRAIL-based cancer therapy in colorectal cancer.
4 . International Symposium on Propionibacteria and Bibfidobacteria
Cork, Ireland

hal-01372131
https://hal.archives-ouvertes.fr/hal-01372131
PRODINRA : 371605

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Recently it has become clear that exposure to xenobiotics may result in various forms of cell death; not only passive cell deaths like necrosis, or programmed cell deaths such as apoptosis, but also regulated necrosis, autophagy, senescence, or mitotic catastrophe. Complex cell signaling networks influence the processing of cell death. Furthermore, recent research has revealed early complex molecular interactions between organelles prior to the final triggering of cell death. The plasma membrane may play an important role in the early cell death signaling events. Regarding this latter aspect, drugs and environmental pollutants have been reported to affect plasma membrane characteristics which may further affect cell fate. Changes in membrane fluidity or in composition and function of specialized membrane microdomains (plasma membrane remodeling) have been proven to be involved in the regulation of many important physiological signaling pathways, including cell death. Furthermore, it has been suggested that a crosstalk between chemical-induced cellular membrane effects and other organelles may be of vital importance to explain the final outcome of chemical exposure. Here, we review the effects of plasma membrane remodeling on cell survival and cell death; we describe how the cell signaling pathways activated by changes in plasma membrane characteristics may influence cell fate. Since plasma membrane function plays an important role in the regulation of a number of cellular responses, it has been implicated in the development or progress of several diseases. A better knowledge of the effects of various chemicals on plasma membrane remodeling may be important for understanding the pathogenesis of major diseases, and may assist in developing new therapeutic strategies.
ISSN: 0300-483X

inserm-00818916
http://www.hal.inserm.fr/inserm-00818916
DOI : 10.1016/j.tox.2012.12.014
PUBMED : 23287390

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Binding of ligand FasL to its receptor Fas triggers apoptosis via the caspase cascade. FasL itself is homotrimeric, and a productive apoptotic signal requires that FasL be oligomerized beyond the homotrimeric state. We generated a series of FasL chimeras by fusing FasL to domains of the Leukemia Inhibitory Factor receptor gp190 which confer homotypic oligomerization, and analyzed the capacity of these soluble chimeras to trigger cell death. We observed that the most efficient FasL chimera, called pFasL, was also the most polymeric, as it reached the size of a dodecamer. Using a cellular model, we investigated the structure-function relationships of the FasL/Fas interactions for our chimeras, and we demonstrated that the Fas-mediated apoptotic signal did not solely rely on ligand-mediated receptor aggregation, but also required a conformational adaptation of the Fas receptor. When injected into mice, pFasL did not trigger liver injury at a dose which displayed anti-tumor activity in a model of human tumor transplanted to immunodeficient animals, suggesting a potential therapeutic use. Therefore, the optimization of the FasL conformation has to be considered for the development of efficient FasL-derived anti-cancer drugs targeting Fas.
ISSN: 1932-6203

hal-00873684
https://hal.archives-ouvertes.fr/hal-00873684
https://hal.archives-ouvertes.fr/hal-00873684/document
https://hal.archives-ouvertes.fr/hal-00873684/file/Functional_Characterization-accepted.pdf
DOI : 10.1371/journal.pone.0054000

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
The implication of oxidative stress as primary mechanism inducing doxorubicin (DOX) cardiotoxicity is still questionable as many in vitro studies implied supra-clinical drug doses or unreliable methodologies for reactive oxygen species (ROS) detection. The aim of this study was to clarify whether oxidative stress is involved in compliance with the conditions of clinical use of DOX, and using reliable tools for ROS detection. We examined the cytotoxic mechanisms of 2 μM DOX 1 day after the beginning of the treatment in differentiated H9c2 rat embryonic cardiac cells. Cells were exposed for 2 or 24 h with DOX to mimic a single chronic dosage or to favor accumulation, respectively. We found that apoptosis was prevalent in cells exposed for a short period with DOX: cells showed typical hallmarks as loss of anchorage ability, mitochondrial hyperpolarization followed by the collapse of mitochondrial activity, and nuclear condensation. Increasing the exposure period favored a shift to necrosis as the cells preferentially exhibited early DNA impairment and nuclear swelling. In either case, measuring the fluorescence lifetime of 1-pyrenebutyric acid or the intensities of dihydroethidium or amplex red showed a consistent pattern in ROS production which was a slight increased level far from representative of an oxidative stress. Moreover, pre-treatment with dexrazoxane provided a cytoprotective effect although it failed to detoxify ROS. Our data support that oxidative stress is unlikely to be the primary mechanism of DOX cardiac toxicity in vitro.
ISSN: 0300-8177

hal-01285736
https://hal-univ-perp.archives-ouvertes.fr/hal-01285736
DOI : 10.1007/s11010-016-2653-x

Éditeur(s) : HAL CCSD Elsevier
Résumé : Thanks to Dr M. Blanchard-Desce (UMR CNRS 5255, University of Bordeaux, France), who kindly provided us with fluorescent dye RS19 and the PIXEL, MRIc, flow cytometry and H2P2 platforms (SFR Biosit, UMS CNRS 3480/US INSERM 018, Rennes).
International audience
[1-9-NαC]-crourorb A1 is a cyclic peptide isolated from Croton urucurana Baillon latex, found in midwestern Brazil, that has been shown to exert cytotoxic effects against a panel of cancer cell lines. However, the underlying mechanisms responsible for the crourorb A1-induced cytotoxicity in cancer cells remain unknown. In this study, the effects of crourorb A1 on the viability, apoptosis, cell cycle and migration of Huh-7 (human hepatocarcinoma) cells were investigated. We evaluated the viability of Huh-7 cells treated with crourorb A1 in 2D and 3D collagen cultures and found that cells in 3D culture exhibited increased resistance to crourorb A1 compared to cells in 2D culture (IC50: 62μg/ml versus 35.75μg/ml). Crourorb A1 treatment decreases the viability of Huh-7 cells in a dose- and time-dependent manner and is associated with the induction of apoptosis, in the absence of necrotic cells, through the activation of caspase-3/7 and increased expression of the pro-apoptotic proteins Bak, Bid, Bax, Puma, Bim, and Bad. The effects of crourorb A1 are also associated with G2/M phase cell cycle arrest and increases in cyclin-dependent kinase (CDK1) and cyclin B1 expression. A significant reduction in Huh-7 cell migration induced by crourorb A1 was also observed in the presence of mitomycin C. Finally, we showed that the JNK/MAP pathway, but not ERK signaling, is involved in crourorb A1-induced hepatocarcinoma cell mortality.
ISSN: 0378-4274

hal-01526429
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01526429
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01526429/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01526429/file/de%20Matos%20C%C3%A2ndido-Bacani%20et%20al.%20-%20%5B1%E2%80%939-N%CE%B1C%5D-crourorb%20A1%20isolated%20from%20Croton%20urucura.pdf
DOI : 10.1016/j.toxlet.2017.03.020

Résumé : La zone latérale des filaments branchiaux des bivalves Codakia orbiculata et Lucina pensylvanica est le lieu d'une symbiose chimioautotrophe avec des bactéries sulfo-oxydantes hébergées dans des cellules spécialisées, appelées bactériocytes. Dans cette étude, nous nous sommes proposés de déterminer les mécanismes qui sous-tendent la plasticité cellulaire et tissulaire observée au cours des processus de décolonisation et de recolonisation bactérienne. Pour ce faire, des individus collectés dans leur milieu naturel, ont été maintenus au laboratoire dans des bacs d'eau de mer filtrée en absence de nourriture et de soufre réduit, afin de provoquer la décolonisation bactérienne des filaments branchiaux. Lorsque la branchie apparaissait purgée de ses symbiotes, les individus ont été remis dans leur habitat naturel afin de provoquer sa recolonisation. L'analyse des branchies au cours de ces processus a fait appel à des techniques variées (histologie, immunohistochimie, hybridation in situ, cytométrie en flux, dosage des protéines et spectrométrie de fluorescence X). Les résultats obtenus ont permis de montrer que l'acquisition environnementale mise en évidence chez les juvéniles des Codakia se poursuivait tout au long de la vie des adultes. Cette étude a également permis une meilleure compréhension des mécanismes tissulaires sous-jacents à la plasticité du filament branchial en mettant en évidence les processus d'apoptose et de prolifération cellulaire qui ont lieu au cours des processus de décolonisation et de recolonisation. Ces processus s'accompagnent d'une variation de la teneur en soufre élémentaire, ainsi que de la taille relative et du contenu génomique des symbiotes
The lateral zone of gills filaments of coastal bivalves Codakia orbiculata and Lucina pensylvanica is the site of chemoautotrophic symbiosis with sulfur-oxidizing bacteria, housed in specialized cells called bacteriocytes. The objective of this thesis is to determine the mechanisms underlying cel1 plasticity and tissue plasticity observed in the lateral zone of gills filaments during the processes of bacterial decolonization and recolonization. In order to do this, the individuals collected in their natural habitat were maintained at the laboratory in seawater filtered tanks, without food and reduced sulfur, to cause bacterial decolonization. When the gills seemed to be purged, the individuals were returned to their natural habitat in order to cause the bacterial recolonization of gills filaments. The analysis of the gills during these processes involves several techniques (histology, immunohistochemistry, molecular hybridization, flow cytometry, total protein assays, protein sulfur assays, X-ray fluorescence spectrometry).This study shows that symbiont acquisition can occur during the entire life of Codakia bivalves. It also allows a better understanding of gills filaments plasticity by highlighting apoptosis and cell proliferation during decolonization and recolonization processes.Theses processes are accompanied with of elemental sufur, relative size and genomic content of symbiontes.

http://www.theses.fr/2011AGUY0461/document

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Apoptosis is a fundamental process that contributes to tissue homeostasis, immune responses, and development. The receptor CD95, also called Fas, is a member of the tumor necrosis factor receptor (TNF-R) superfamily. Its cognate ligand, CD95L, is implicated in immune homeostasis and immune surveillance, and various lineages of malignant cells exhibit loss-of-function mutations in this pathway; therefore, CD95 was initially classified as a tumor suppressor gene. However, more recent data indicate that in different pathophysiological contexts, this receptor can transmit non-apoptotic signals, promote inflammation, and contribute to carcinogenesis. A comparison with the initial molecular events of the TNF-R signaling pathway leading to non-apoptotic, apoptotic, and necrotic pathways reveals that CD95 is probably using different molecular mechanisms to transmit its non-apoptotic signals (NF-κB, MAPK, and PI3K). As discussed in this review, the molecular process by which the receptor switches from an apoptotic function to an inflammatory role is unknown. More importantly, the biological functions of these signals remain elusive.
ISSN: 0006-3002

hal-01061290
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01061290
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01061290/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01061290/file/BBAreviewsoncancer-legembre-final.pdf
DOI : 10.1016/j.bbcan.2014.04.007

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
The shallow-water bivalve Codakia orbiculata which harbors gill-endosymbiotic sulfur-oxidizing ?-proteobacteria can lose and acquire its endosymbionts throughout its life. Long-term starvation and recolonization experiments led to changes in the organization of cells in the lateral zone of gill filaments. This plasticity is linked to the presence or absence of gill-endosymbionts. Herein, we propose that this reorganization can be explained by three hypotheses: (a) a variation in the number of bacteriocytes and granule cells due to proliferation or apoptosis processes, (b) a variation of the volume of these two cell types without modification in the number, and (c) a combination of both number and cell volume variation. To test these hypotheses, we analyzed cell reorganization in terms of proliferation and apoptosis in adults submitted to starvation and returned to the field using catalyzed reporter deposition fluorescence in situ hybridization, immunohistochemistry, and structural analyses. We observed that cell and tissue reorganization in gills filaments is due to a variation in cell relative abundance that maybe associated with a variation in cell apparent volume and depends on the environment. In fact, bacteriocytes mostly multiply in freshly collected and newly recolonized individuals, and excess bacteriocytes are eliminated in later recolonization stages. We highlight that host tissue regeneration in gill filaments of this symbiotic bivalve can occur by both replication of existing cells and division of undifferentiated cells localized in tissular bridges, which might be a tissue-specific multipotent stem cell zone.
ISSN: 1059-910X

hal-01545915
https://hal.archives-ouvertes.fr/hal-01545915
DOI : 10.1002/jemt.22041

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
Cnidarian-dinoflagellate symbiosis disruption and subsequent bleaching are major concerns, especially regarding their ecological consequences on coral reefs and temperate coralligenous communities. Cnidarian bleaching is caused by a variety of environmental stressors, such as elevated seawater temperature associated with global climate change, and by pollutants, such as herbicides and metals. Several cellular events have been described to explain symbiosis dysfunction and bleaching. Excess or damaged Symbiodinium symbionts are removed through a variety of mechanisms, including exocytosis, apoptosis, necrosis and autophagy. However, few studies have compared in the same species the relative involvement of these mechanisms, according to the stress inducing the bleaching. In this study, we used two different treatments —temperature and menthol— to induce bleaching in the sea anemone Anemonia viridis. By monitoring the ultrastructural tissue modifications, in control specimens we observed a basal rate of in situ symbiont digestion —or symbiophagy— induced by starvation. Symbiophagy was strongly induced in menthol-treated specimens and was the main cellular process of bleaching, whereas apoptosis and necrosis predominated in hyperthermal-induced bleaching. These results suggested a host effect through autophagy in menthol-treated specimens. These observations also suggested that symbiont removal may result from reengagement of the phagosomal maturation process in the host. These overall data demonstrate that several Symbiodinium cell removal mechanisms coexist and that stressors can activate one or more of these pathways, depending on the stress type, intensity or duration.
ISSN: 0334-5114

hal-01546908
http://hal.upmc.fr/hal-01546908
DOI : 10.1007/s13199-016-0406-y

Éditeur(s) : HAL CCSD
Résumé : The ovary is not solely the unique source of oocytes necessary for fertilization but also a crucial conductor of endocrine regulations of the reproductive system. In the fetal ovary, oocytes grouped in clusters are surrounded by somatic pre-granulosa cells, altogether delineated by a basement membrane and constituting ovarian cords. The formation of the definitive ovarian functional units requires their fragmentation into follicles formed by a single oocyte surrounded by granulosa cells and delineated by a basement membrane. This partitioning occurs within a tiny period and is dependent on the correct timing of the meiotic process, on an apoptotic wave that specifically targets oocytes and the remodelling of the basement membrane. Follicle formation is a crucial event of ovarian development because the follicle stock formed at the end of this process is non-renewable. Therefore, any disturbance of this process can lead to reproductive abnormalities in adulthood. Recent data on endocrine disruptors displaying estrogenic activity have involved the fragile estrogenic homeostasis in the process. We aimed at better understanding how the endogen estrogen 17b-estradiol (E2) contributes to follicle formation. To assess it, Sprague-Dawley rats were treated with E2 at different doses during follicle formation, /i.e./ in the 3 days following birth. We show that although this treatment impairs the development of the hepatic and ovarian detoxification systems, the female neonate is able to increase its estrogen clearance capabilities. Nevertheless, E2 treatment within this critical period triggers a dose-dependent decrease of oocyte number per ovary. Regardless of the E2 dose, puberty is advanced. All treated females are at least transiently fertile, yet these reproductive capabilities rapidly decline. A high (10 µg/day) dose of E2 leads to a secondary infertility characterized by anovulation that probably result from a dysfunction of the whole reproductive tract. By contrast, a lower (0.1 µg/day) dose of E2, that does not significantly modify neonatal oocyte survival, leads to a progressive reproductive senescence likely due to ovarian failure. Long-term troubles may originate from precocious E2 impact on the ovary since it disturbs ovarian transcriptome and produces many lesions on ovarian cell DNA.
L'ovaire est au cœur de la physiologie de la reproduction féminine. Il est à la fois à l'origine des ovocytes nécessaires à la fécondation et acteur des régulations endocrines du système reproducteur. Les ovocytes de l'ovaire fœtal sont groupés en amas bordés de cellules somatiques et d'une membrane basale, formant les cordons ovariens. Ces cordons ovariens se fragmentent pour former les follicules ovariens: un ovocyte bordé de cellules somatiques et d'une membrane basale. Cette fragmentation se déroule sur une courte période (/in utero/ chez la femme et juste après la naissance chez les rongeurs) et fait intervenir une vague de mort cellulaire programmée touchant les ovocytes pendant leur méiose et un remodelage de la membrane basale. La formation des follicules est une période cruciale du développement ovarien puisque le stock de follicules formés à l'issue de ce processus est non renouvelable. Son altération peut donc être à l'origine de troubles de la fonction de reproduction chez le futur adulte. Il existe une fragile homéostasie œstrogénique en place au moment de la formation folliculaire: chez la femme, l'ovaire se développe /in utero/, sous l'influence des œstrogènes circulants maternels et la production ovarienne fœtale. Chez les rongeurs, les follicules se forment lors de la levée de l'imprégnation hormonale maternelle au moment de la naissance. Nous avons souhaité comprendre comment le 17b-œstradiol (E2), œstrogène endogène, pouvait contribuer à la mise en place des follicules. Pour cela, des rattes Sprague-Dawley ont été traitées pendant la période de formation des follicules avec de l'E2 à différentes doses entre 0.01 et 10 µg/jour. Nos travaux démontrent que le traitement de rattes par des œstrogènes entre leur naissance et 3 jours induit une réduction dose-dépendante de leur nombre d'ovocytes. Bien que le traitement entrave le développement normal du système de détoxication hépatique et ovarien, l'animal est capable d'accroître ses capacités d'élimination des œstrogènes. Ces capacités sont toutefois insuffisantes pour bloquer les effets du traitement. Quelle que soit la dose d'E2 utilisée (0.1 ou 10 µg/jour), la puberté est plus précoce chez les femelles exposées mais toutes sont fertiles, au moins transitoirement. Cependant le traitement induit une dégradation rapide des capacités reproductives. L'E2 modifie la dynamique folliculaire chez l'adulte, longtemps après l'arrêt du traitement. Si une forte dose d'E2 réduit la survie des ovocytes lors de la formation des follicules et conduit à une infertilité secondaire due à un dysfonctionnement général du tractus reproducteur, une dose faible n'affecte pas la survie des ovocytes à court terme mais conduit à une sénescence reproductive précoce vraisemblablement d'origine ovarienne. L'origine de ces troubles pourrait résider dans les effets précoces de l'exposition à E2 : cette molécule altère le transcriptome ovarien et induit de nombreuses lésions de l'ADN des cellules ovariennes.
https://tel.archives-ouvertes.fr/tel-00953657

NNT : 2013REN1S107
tel-00953657
https://tel.archives-ouvertes.fr/tel-00953657
https://tel.archives-ouvertes.fr/tel-00953657/document
https://tel.archives-ouvertes.fr/tel-00953657/file/CHALMEY_Clementine.pdf

Éditeur(s) : HAL CCSD
Résumé : International audience
Fucoxanthin is a carotenoid present in brown micro-and macro algae, that induces apoptosis or autophagy in cancer cells grown in vitro. In vivo studies confirmed its interest as a natural anticancer compound, as it exerts antitumoral, antimetastatic and antiangiogenic activities in animal models. Studies focused on the pharmacology of fucoxanthin in cancer cells and tumors have revealed that it affects a wide panel of cellular, molecular and tissular processes, suggesting that its biological activity may be related in part to a nonspecific integration in cell membranes, and possible interaction with lipid rafts. Thus, preliminary data confirming this interaction of fucoxanthin with lipid rafts were obtained in mast cells and hepatoma WIF-B9 cells. We here discuss this hypothesis, in view of the critical function of lipid rafts in cancer cell survival, invasivity and communication with the tumoral microenvironment.
ISSN: 2141-2294

hal-01391001
https://hal.archives-ouvertes.fr/hal-01391001
https://hal.archives-ouvertes.fr/hal-01391001/document
https://hal.archives-ouvertes.fr/hal-01391001/file/could-fucoxanthin-interaction-with-lipid-rafts-mediate-its-cytotoxicity-incancer-cells-.pdf
DOI : 10.4172/jomr.1000144

Éditeur(s) : HAL CCSD
Résumé : Staphylococcus aureus is an opportunistic pathogen of humans and warm-blooded animals. It is foundin healthy carriage and can cause a wide range of infections in its hosts. The epithelial surface is theprincipal site of staphylococcal infection. The pathogens have evolved sophisticated mechanisms tohijack the main function of the host cells, thus promoting their invasion and colonization. Amongstthese effects are, alteration of host cell apoptosis, promotion of cell proliferation or conversely,inhibition of cell growth. Using human cervix cancer HeLa and mammary gland bovine MAC-Tepithelial cells, we studied the cytopathic effect of S. aureus and its ability to affect host cell cycle.Using microscopic examination, we found that S. aureus induced the cytopathic effect resulting in theenlargement of the host cells 72 hours post infection. Our experiments showed that the proliferationrate of the epithelial cells was slowed down in S. aureus-infected culture. Three different S. aureusstrains from human and animal origins were used and showed similar effect on both human and bovinecells. Microscopic analysis of DAPI-stained cells revealed huge decrease of mitotic indexes in infectedHeLa and Mac-T cells, synchronized by Double Thymidine Block. Using flow cytometry analysis, wefound that S. aureus induces G2/M phase delay in synchronous HeLa cells. By Western blot analysis,we showed that the G2/M phase delay was associated with accumulation of inactive phosphorylatedcyclin-dependent kinase Cdk1/cdc2, a key inducer of mitosis entry, and dephosphorylation of corehistone H3, an important component of the epigenetic landscape.These results suggest that the delay in G2/M phase transition of the infected epithelial cells may beone of the mechanisms employed by S. aureus to survive and to propagate inside the host. A fullcomprehension of the mechanisms associated to S. aureus-host cell interactions will undoubtedly helpto discover novel targets to treat staphylococcal infections.
EMBO
Villars sur Ollon, Switzerland

hal-01454214
https://hal.archives-ouvertes.fr/hal-01454214
PRODINRA : 171657

Éditeur(s) : HAL CCSD
Résumé : International audience
Necroptosis is a regulated form of cell death involved in several disease models including in particular liver diseases. Receptor-interacting protein kinases, RIPK1 and RIPK3, are the main serine/threonine kinases driving this cell death pathway. We screened a noncommercial, kinase-focused chemical library which allowed us to identify Sibiriline as a new inhibitor of necroptosis induced by tumor necrosis factor (TNF) in Fas-associated protein with death domain (FADD)-deficient Jurkat cells. Moreover, Sib inhibits necroptotic cell death induced by various death ligands in human or mouse cells while not protecting from caspase-dependent apoptosis. By using competition binding assay and recombinant kinase assays, we demonstrated that Sib is a rather specific competitive RIPK1 inhibitor. Molecular docking analysis shows that Sib is trapped closed to human RIPK1 adenosine triphosphate-binding site in a relatively hydrophobic pocket locking RIPK1 in an inactive conformation. In agreement with its RIPK1 inhibitory property, Sib inhibits both TNF-induced RIPK1-dependent necroptosis and RIPK1-dependent apoptosis. Finally, Sib protects mice from concanavalin A-induced hepatitis. These results reveal the small-molecule Sib as a new RIPK1 inhibitor potentially of interest for the treatment of immune-dependent hepatitis. © 2017 Federation of European Biochemical Societies
ISSN: 1742464X

hal-01616420
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01616420
DOI : 10.1111/febs.14176
PUBMED : 28715128

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Many studies have reported proliferative, differentiating or protective effects of estradiol, notably through estrogen receptor alpha (ERα). On the contrary, the ligand-independent action of ERα is currently poorly documented notably in cell protection. The stable transfection of wild type, substituted or truncated form of ERα in PC12 cells (ERα negative cell line) lead the specific study of its ligand-independent action. Hence, we demonstrate here that, in the absence of E2, the expression of ERα prevents cells from apoptosis induced by serum deprivation. This protection is not due to an ERE-mediated transcription and does not require either AF-1 or AF-2 transactivation functions. It is afforded to the Y537 residue of ERα and activation of c-Src/Stat3 signaling pathway.
ISSN: 1932-6203

inserm-00865332
http://www.hal.inserm.fr/inserm-00865332
http://www.hal.inserm.fr/inserm-00865332/document
http://www.hal.inserm.fr/inserm-00865332/file/PMID_23825704.pdf
DOI : 10.1371/journal.pone.0069081

Éditeur(s) : HAL CCSD Wiley-Blackwell
Résumé : International audience
The shallow-water bivalve Codakia orbiculata which harbors gill-endosymbiotic sulfur-oxidizing g-proteobacteria can lose and acquire its endosymbionts throughout its life. Longterm starvation and recolonization experiments led to changes in the organization of cells in the lateral zone of gill filaments. This plasticity is linked to the presence or absence of gill-endosymbionts. Herein, we propose that this reorganization can be explained by three hypotheses: (a) a variation in the number of bacteriocytes and granule cells due to proliferation or apoptosis processes, (b) a variation of the volume of these two cell types without modification in the number, and (c) a combination of both number and cell volume variation. To test these hypotheses, we analyzed cell reorganization in terms of proliferation and apoptosis in adults submitted to starvation and returned to the field using catalyzed reporter deposition fluorescence in situ hybridization, immunohistochemistry, and structural analyses. We observed that cell and tissue reorganization in gills filaments is due to a variation in cell relative abundance that maybe associated with a variation in cell apparent volume and depends on the environment. In fact, bacteriocytes mostly multiply in freshly collected and newly recolonized individuals, and excess bacteriocytes are eliminated in later recolonization stages. We highlight that host tissue regeneration in gill filaments of this symbiotic bivalve can occur by both replication of existing cells and division of undifferentiated cells localized in tissular bridges, which might be a tissue-specific multipotent stem cell zone.
Microscopy Research and Technique

hal-00743036
https://hal-uag.archives-ouvertes.fr/hal-00743036

Éditeur(s) : HAL CCSD Oxford University Press (OUP)
Résumé : International audience
BACKGROUND: Phthalic acid esters are widely used in the manufacture of plastics. Numerous studies have shown that these phthalates impair testicular testosterone production in the rat. However, the scarce and contradictory data concerning humans have cast doubt over whether these compounds are also anti-androgenic in man. We therefore investigated the direct effects of di-(2-ethylhexyl) phthalate (DEHP) and mono-(2-ethylhexyl) phthalate (MEHP) on organo-cultured adult human testis and a human cell line. METHODS: Adult human testis explants or NCI-H295R adrenocortical human cells were cultured with DEHP or MEHP. The effects of ketoconazole, used as a reference molecule, were also assessed. RESULTS: In both models, DEHP and MEHP significantly inhibited testosterone production. The effects of both phthalates appeared to be specific for steroidogenesis, as INSL3 production by Leydig cells was not altered. Furthermore, the phthalates of interest had no effect on inhibin B production by Sertoli cells or on germ cell apoptosis. As only a small fraction of the phthalates added was found in the testis explants, and as these compounds were found to be metabolized, we estimate that the anti-androgenic effects observed occurred at concentrations of phthalates that are of the same order of magnitude as exposures reported in the literature for men. CONCLUSIONS: We provide the first evidence that DEHP and MEHP can inhibit testosterone production in the adult human testis. This is consistent with recent epidemiological findings of an inverse correlation between exposure to MEHP and testosterone concentrations.
ISSN: 0268-1161

hal-00876965
https://hal.archives-ouvertes.fr/hal-00876965
DOI : 10.1093/humrep/des069
PUBMED : 22402212

Éditeur(s) : HAL CCSD Cancer Research UK
Résumé : International audience
BACKGROUND: Cancer cell killing might be achieved by the combined use of available drugs. Statins are major anti-hypercholesterolemia drugs, which also trigger apoptosis of many cancer cell types, while docetaxel is a potent microtubule-stabilising agent. METHODS: Here, we looked at the combined effects of lovastatin and docetaxel in cancer cells. RESULTS: Whole transcriptome microarrays in HGT-1 gastric cancer cells demonstrated that lovastatin strongly suppressed expression of genes involved in cell division, while docetaxel had very little transcriptional effects. Both drugs triggered apoptosis, and their combination was more than additive. A marked rise in the cell-cycle inhibitor p21, together with reduction of aurora kinases A and B, cyclins B1 and D1 proteins was induced by lovastatin alone or in combination with docetaxel. The drug treatments induced the proteolytic cleavage of procaspase-3, a drop of the anti-apoptotic Mcl-1 protein, Poly-ADP-Ribose Polymerase and Bax. Strikingly, docetaxel-resistant HGT-1 cell derivatives overexpressing the MDR-1 gene were much more sensitive to lovastatin than docetaxel-sensitive cells. CONCLUSION: These results suggest that the association of lovastatin and docetaxel, or lovastatin alone, shows promise as plausible anticancer strategies, either as a direct therapeutic approach or following acquired P-glycoprotein-dependent resistance.
ISSN: 0007-0920

hal-00684528
https://hal.archives-ouvertes.fr/hal-00684528
DOI : 10.1038/bjc.2012.6
PUBMED : 22294184

Éditeur(s) : HAL CCSD Nature Publishing Group
Résumé : International audience
Hepatocyte death is a central event during liver disease progression, in which immune cells play key roles by activating members of the Tumor Necrosis Factor Receptor Superfamily (TNFRSF), including TNFR1 (TNFRSF1A), Fas (TNFRSF6) and TRAIL-R2 (TNFRSF10B). Receptor Interacting Protein Kinase 1 (RIPK1) emerged as a signaling node downstream of these receptors. In the case of TNFR1, RIPK1 has been demonstrated to paradoxically serve as a scaffold to promote the survival of hepatocytes and as a kinase to kill them. To evaluate whether RIPK1 also protects hepatocytes from death in response to FasL or TRAIL, we took advantage of liver parenchymal cell-specific Ripk1 knockout mice (Ripk1 (LPC-KO)). We found that Ripk1 (LPC-KO) mice, as well as primary hepatocytes derived from them, were more susceptible to Fas-mediated apoptosis than their respective WT counterparts. Fas-induced hepatocyte death was independent of TNF-α signaling. Interestingly, while TRAIL administration did not induce hepatitis in Ripk1 (LPC-KO) mice or in their WT counterparts, its combination with IFN-γ only induced TNF-α dependent apoptosis in the Ripk1 (LPC-KO) mice. Together, our data demonstrate the protective role of RIPK1 downstream of Fas and highlight the general protective function of RIPK1 in hepatocytes exposed to inflammatory conditions, where TNF-α, FasL and/or TRAIL are present.
ISSN: 2045-2322

hal-01579612
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01579612
DOI : 10.1038/s41598-017-09789-8
PUBMED : 28835677