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Etude des réponses du métabolisme de Dunaliella sp au stress abiotique pour une exploitation en thérapie : analyses génomiques et protéomiques

Authors: Elleuch, Fatma;

Etude des réponses du métabolisme de Dunaliella sp au stress abiotique pour une exploitation en thérapie : analyses génomiques et protéomiques

Abstract

In this work, we optimized the culture conditions of Dunaliella salina sp, isolated from the Tunisian coast to increase carotenoids production. For that, we performed a Box-Behnken experimental design to study the effects and interactions between salinity, light intensity and nitrogen supply conditions. We succeeded to improve the intracellular production of carotenoids by five-fold. Subsequently, we studied the characteristics of cultured cells under stress (DSS) and control (DSC) conditions as well as the transcriptional modifications of the genes involved in stress response and β-carotene synthesis. Amongst studied genes, Lycopene β cyclase gene (LCYB) and Carotene globular protein gene (Cgp) were significantly overexpressed. 3D structures of the corresponding proteins and their posttranslational modifications have been determined. The anticancer activity of this strain cultivated under normal (DSC) or stressed (DSS) conditions was explored showing that the aqueous extract of DSS was four times more toxic than DSC one against murine breast cancer 4T1 cells. The extract was also active against A549 human lung adenocarcinoma cells and human Hela cells of cervical cancer. The study of the genomic DNA fragmentation of the treated 4T1 cells indicates that the extract induces an apoptosis cell death. Validation of these results by Western Blot was also performed by proving the activation of caspases 3 and 9 and the deactivation of PARP-1. In vivo experiments were performed on Balb/c mice that have been orthotopically implanted with 4T1 cells into the mammary glands. The results show a delay of the tumor growth and a decrease of the tumor size in mice treated with DSS aqueous extract. Interestingly, an activation of the anti-cancer immunity was observed with an increase of Natural Killer cells, activated cytotoxic lymphocytes and plasmacytoid DC as well as repression of immunosuppressive myeloid cells and enzymes (Arg-1, NOS-2 and COX-2). Altogether, our data reveal the potentiality of D. salina aqueous extract grown under stressed conditions as anti-cancer agent, which deserves attention in the future.

Nous nous sommes intéressés à optimiser la production de caroténoïdes par les microalgues Dunaliella salina isolées des côtes tunisiennes, via un plan d’expérience du type Box-Behnken en étudiant les effets et les interactions entre la salinité, l’intensité lumineuse et l’approvisionnement en azote. Nous avons trouvé des conditions permettant une augmentation de 5 fois la production intracellulaire de caroténoïdes. Nous avons étudié les caractéristiques des cellules cultivées en condition de stress (DSS) ou non (DSC) et les modifications transcriptionnelles des gènes impliqués dans la réponse au stress et la synthèse de β-carotène. Les gènes de la Lycopène β cyclase (LCYB) et de la protéine globulaire associée au carotène (Cgp) sont surexprimés. Les structures 3D des deux protéines codées par ces gènes et leurs modifications post-traductionnelles ont été déterminées. L’activité anticancéreuse d’extrait de cette souche cultivée dans des conditions normales (DSC) ou stressées (DSS) a été testée sur des cellules 4T1 de cancer de sein murin. L’extrait aqueux DSS est plus toxique que celui issu de DSC. L’étude de la fragmentation de l’ADN génomique des cellules traitées met en évidence une apoptose avec une activation des caspases 3 et 9 et la désactivation de PARP-1. Pour valider ces résultats, une expérimentation in vivo a été menée sur des souris Balb/c implantées avec des cellules 4T1 dans la glande mammaire. Les résultats montrent une diminution de la taille de la tumeur des souris traitées et un retard de croissance. Cette activité a été accompagnée d’une activation de l’immunité anticancéreuse chez les souris traitées avec une infiltration de cellules Natural killers, de lymphocytes cytotoxiques et de cellules dendritiques plasmacytoÏdes ainsi qu’une diminution de macrophages immunosuppresseurs. L’ensemble de nos résultats révèle la potentialité de l’extrait aqueux de cette micro algue comme anti-cancéreux qui mériterait d’être développée ultérieurement.

Keywords

Dunaliella, [SDV.BIBS] Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Apoptose, Molecular modeling, Apoptosis, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Réponse immunitaire, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], [SDV.BC.BC] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Étude transcriptionnelle, Immune response, Modélisation moléculaire, Transcriptional study, Cancer

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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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