A qualidade do ar nas áreas urbanas tende a apresentar concentrações indesejáveis de contaminantes provenientes da queima de biomassa, não havendo um sistema abrangente de monitoramento. Achados in vivo mostram que o diesel pode conter compostos que modulam a atividade do estrógeno. Estudos in vitro apoiam essa ideia, como o estudo realizado com cloreto de metileno extraído do diesel que comprova atuar como ativadores de ligação para os receptores de estrógeno. Esse trabalho tem por objetivo avaliar o efeito da exposição à metais presentes nas partículas de exaustao de diesel (PED) na morfologia do ovaário através da histologia, e análise do padrão de expressão dos receptores de estrógeno (ERs) no ovário de zebrafish (Danio rerio) usando a metodologia de PCR em tempo real. Fêmeas adultas de zebrafish foram expostas à concentrações ambientais de combinado de metais (Ni = 0,181 mg/L ; Fe = 0,07455 mg/L ; Pb= 0,05 mg/ L; Cd = 0,029 mg/ L; Cr = 0,161 mg/L ; Cu = 0,017 mg / L) por 3 dias. Em relação a contagem de ovócitos atrésicos a média observada nos animais expostos foi de 1,5 ± 0,4 e nos controles foi de 1,34 ± 0,3. Contudo, não houve diferença significativa entre os grupos (p > 0,05). Na expressão dos três genes receptores de estrógeno (esr1, esr2a, esr2b) as médias foram: 0,68; 1,32; 0,54; respectivamente. No entanto, não houve alteração significativa na expressão desses genes em relação aos controles. Os elementos traços presentes nas PED em concentração ambiental e em exposição aguda na fase adulta não alteram a expressão dos ERs o também não afetam o número de folículos atrésicos nos ovários / The air quality in urban areas tends to have undesirable concentrations of contaminants from biomass combustion, without a monitoring system. In vivo findings show that diesel may contain compounds that modulate the activity of estrogen. In vitro studies support this idea, as the study with methylene chloride extract from diesel proves that this substance act as activators binding to the estrogen receptors (ERs). This study aims to evaluate the effect of exposure to metals present in diesel exhaust particles (PED) in ovary morphology through histology, and analysis of expression pattern of ERs in ovary of zebrafish (Danio rerio) using real time PCR metodology. Adult females were exposed to ambient concentrations of combined metals (Ni = 0.181 mg / L, Fe = 0.07455 mg / L Pb = 0,05 mg / L; Cd = 0.029 mg / L; = 0.161 mg Cr / L, Cu = 0.017 mg / L) for 3 days. The mean of atretic oocytes counting observed in animals exposed was 1.5 ± 0.4 and the controls was 1.34 ± 0.3. However, no significant difference between groups was observed (p> 0.05). The expression of the three genes estrogen receptor (esr1, esr2a, esr2b) mean were: 0.68, 1.32, 0.54, respectively. However, no significant change in the expression of these genes in relation to controls was observed. In conclusion, the trace elements present in the PED concentration in environmental and acute exposure in adulthood not alter the expression of ERs and does not affect the number of atretic follicles in the ovary

La résistance à l’hormonothérapie est un challenge majeur en clinique dans le choix du traitement chez les patientes atteintes de cancer du sein RE+ et l’activation des voies de signalisation PI3K/Akt/mTOR ou MAPK semble être impliquée dans la résistance à l’hormonothérapie. L’objectif de ce travail a été dédié à l’exploration de stratégies de réversion de la résistance à l’hormonothérapie et à l’identification de nouveaux biomarqueurs associés à cette résistance. Nous avons identifié dans des lignées résistantes à l’hormonothérapie ayant acquis l’activation endogène des voies PI3K/Akt/mTOR et MAPK, l’activation aberrante du système ErbB, pouvant être à la base de l’activation de ces deux voies de signalisation. L’inhibition d’au moins une des deux voies (par un inhibiteur de mTOR, un inhibiteur de PI3K et/ou un inhibiteur de MEK) a conduit dans la lignée sensible à une augmentation de la sensibilité au tamoxifène et au fulvestrant et dans les lignées résistantes à une restauration de la sensibilité à l’hormonothérapie. La réversion de la résistance au fulvestrant par la rapamycine a été démontrée non seulement au niveau de la prolifération cellulaire mais aussi au niveau de l’expression des gènes, explorés par une approche génomique. Par ailleurs, par une approche gènes candidats, nous avons identifié une signature de trois gènes (TACC1, NOV et PTTG1) présentant une valeur pronostique et associée à la résistance à l’hormonothérapie pouvant représenter un nouvel outil de diagnostic des patientes atteintes de cancer du sein hormono-dépendant. / Endocrine therapy resistance is one of the main challenges in the treatment of estrogen receptor positive breast cancer patients and the activation of PI3K/Akt/mTOR or MAPK signaling pathways seems to be implicated in the acquisition of the resistance. The objective of this work has been dedicated to the exploration of strategies to reverse the resistance to endocrine therapy and the identification of new biomarkers associated with this resistance. We have identified in cellular models resistant to hormone therapy, which have acquired the endogenous activation of PI3K/Akt/mTOR and MAPK pathways, the aberrant activation of the ErbB system which may be the cause of the activation of these pathways. The inhibition of at least one of these two pathways (by an mTOR inhibitor), a PI3K inhibitor) and/or a MEK inhibitor) was shown to increase sensitivity to tamoxifen and fulvestrant in the sensitive cells and to restore sensitivity to endocrine therapy in the resistant cells. The reversion of resistance to fulvestrant by rapamycin has been demonstrated not only at the cell proliferation level but also at the gene expression level explored by a genomic approach. In addition, using a candidate gene approach, we identified a signature of three genes (TACC1, NOV and PTTG1) with prognostic value and associated with resistance to endocrine therapy. These genes may represent a new diagnostic tool for patients treated with endocrine therapy.

Purpose: to evaluate if there is a difference between adjuvant chemo-endocrine therapy (QHT) and hormone therapy (HT) alone in ten years overall survival (OS) in post-menopausal women with early stage luminal A breast cancer Methods: A non concurrent cohort study was conducted in a cancer treatment center in Peru, we measured demographic and clinical-pathologic anatomy variables. Log-rank test and a Kaplan-Meier (KM) curve were performed to evaluate ten years OS. Cox regression analysis was used and hazard ratio were reported with confidence intervals 95% (95%CI) for crude and adjusted by the significant variables in the bivariate analysis. The fullfilment of hazard proportionality was evaluated by Schoenfeld residuals method and graphic method. Results: 65 patients received adjuvant chemo-endocrine therapy and 140 only received hormone therapy. Ten years OS was 77% for QHT and 84% for HT, this difference was not significant when using KM and log-rank; age at diagnosis (p=0,01), clinical status (p=0,02), tumor size (p=0,04), positive estrogen receptor (p=0,03), node status (p=0,012) and type of surgery (p=0,03) were statistically significant when compare with OS. When proportional hazards assumption was evaluated (SPH), only the period of time after two years of following was satisfied, cox models were created for this period of time. Crude HR for ten years OS was 1,48 (CI95%:0,65-3,39). First model adjusted HR was 1,83 (CI95%:0,64-5,30) and second model adjusted HR was 1,77 (CI95%:0,64-4,90). Conclusions: There was no significant difference in ten years OS between both courses of treatment evaluated in post-menopausal women with luminal A breast cancer. / Objetivo: evaluar si existe diferencia en la supervivencia global (SG) a diez años entre la quimioterapia adyuvante asociada a hormonoterapia (QHT) frente a la hormonoterapia sola en mujeres posmenopáusicas diagnosticadas con cáncer de mama luminal A (CMLA) en estadio temprano. Métodos: se realizó un estudio cohortes no concurrente en un centro de atención oncológica en Perú. Se incluyeron variables demográficas y clínico-patológicas. Para comparar la SG se utilizó la curva de Kaplan-Meier (KM), test de log-Rank y la regresión de Cox para estimar el Hazard Ratio (HR) con intervalos de confianza 95% (IC95%) tanto crudos como ajustados por las variables asociadas durante el análisis bivariado. Se evaluó el cumplimiento del supuesto de proporcionalidad de hazard (SPH) con el método de residuos de Schoenfeld y método gráfico. Resultados: 65 pacientes recibieron QHT y 140 sólo hormonoterapia. La SG a los diez años fue 77% y 84% para QHT y HT respectivamente, esta diferencia no fue significativa al utilizar KM y test de log-Rank; no obstante la edad (p=0,01), estadio clínico (p=0,02), tamaño tumoral (p=0,04), receptor estrogénico positivo (p=0,03), número de ganglios (p=0,012) y tipo de cirugía (p=0,03) resultaron asociadas significativamente a la supervivencia global a los diez años. Cuando se evaluó el SPH se evidenció que sólo se cumplía tras los dos años de seguimiento, por lo que se generaron modelos de Cox en éste periodo. El HR crudo a los diez años fue de 1,48 (IC95%: 0,65-3,39). En el modelo ajustado uno se observó un HR de 1,83 (IC95%: 0,64-5,30) y para el segundo modelo ajustado un HR de 1,77 (IC 95%: 0,64-4,90). Conclusiones: no se encontró diferencia significativa en la SG a los diez años entre los esquemas terapéuticos evaluados en mujeres posmenopáusicas con CMLA.

Parallèlement à leur action nucléaire, les œstrogènes exercent également des effets via une signalisation cytoplasmique par des mécanismes pas complètement élucidés. Nous avons mis en évidence la méthylation de ERα (mERα) sur arginine est l’évènement clé de la signalisation non génomique des œstrogènes dans les cellules tumorales mammaires. Cette méthylation entraîne la formation d’un complexe contenant ERα/SRc/PI3K/FAK qui active des cascades de phosphorylation régulant la prolifération cellulaire. La production d’un anticorps spécifique de la forme méthylée a permis de montrer que ERα est hyperméthylé dans 55% des tumeurs mammaires. Afin de comprendre les mécanismes de régulation de la méthylation, nous avons recherché de nouveaux partenaires impliqués dans ce processus. Au cours de ma thèse, j’ai montré que la protéine kinase LKB1 est impliquée dans la signalisation non génomique des œstrogènes. Dans les cellules MCF-7, les œstrogènes entraînent rapidement le recrutement de LKB1 au sein du complexe précédemment décrit. De plus, LKB1 est indispensable à la formation de ce macro complexe ainsi qu’à la phosphorylation de Bad en aval. En effet, par cette action, LKB1 participe au rôle protecteur des œstrogènes contre l’apoptose orchestré par les œstrogènes. De plus, une étude de l’expression de mERα et LKB1, sur une série de tumeurs mammaires, a montré une corrélation significative entre l’expression de ces deux protéines associée à l’envahissement ganglionnaire. Ces résultats révèlent une signification biologique de l’interaction LKB1/mERα, suggérant un rôle oncogénique putatif de LKB1 / Besides its nuclear action, estrogens mediate also cytoplasmic signaling, however the mechanisms are not fully understood. We recently showed that arginine methylation of estrogen receptor alpha (Erα) is required for the recruitment of PI3K and Src, activating downstream kinases. An antibody that specifically recognized Erα dimethylated was generated allowing the detection of Erα hypermethylation in 55% of breast tumors. To decipher the molecular mechanisms that regulate Erα methylation in non genomic pathways, we investigated new partners of the methylated form of Erα (mERα). We identified the tumor suppressor LKB1, a Ser/Thr kinase involved in cell metabolism and cell polarity as a new partner of mERα. To ascribe a biological role to this interaction, we analyzed the protein complexes containing mErα and LKB1. LKB1 is part of the complexe involved in Erα non genomic pathway. LKB1 is essential for Erα methylation and the formation of the macrocomplex Erα/p85/Src suggesting a functional role of LKB1 in Erα methylation and then activation of downstream signaling pathways. Using a phosphospecific antibody microarray, we observed that LKB1 was required for Bad phosphorylation, suggesting its involvement in apoptosis. Indeed, we found that LKB1 participes in the protective role of estrogen against apoptosis. Interestingly, an IHC study on human breast tumors points a correlation between the expression of LKB1 and mErα : their expression is correlated with lymph node metastasis. Altogether, these results reveal biological significance of mErα/LKB1 interaction, suggesting a putative oncogenic role to LKB1

Le cancer du sein représente une cause de mortalité élevée chez la femme. Le cancer du sein est un cancer hormono-dépendant. De ce fait, il est extrêmement important de définir le rôle joué par les différents acteurs protéiques de la signalisation hormonale, notamment la signalisation œstrogénique. Parallèlement aux effets nucléaires de ERa où l'hormone lie le récepteur nucléaire et régule la transcription génique, il existe une voie dite non génomique. L'équipe a montré que les œstrogènes induisent la méthylation de ERa, qui est un prérequis au recrutement de la Pl3K et de la tyrosine kinase Src, conduisant à l'activation de molécules de signalisation telles que les MAPK et Akt, induisant prolifération et survie cellulaire. Durant ma thèse, j'ai pu démontrer que le complexe mERa/Src/Pl3K existe in vivo et constitue un nouveau biomarqueur indépendant de mauvais pronostique. La recherche de nouveaux partenaires du complexe mERa/Src/Pl3K nous a permis d'identifier le suppresseur de tumeur LKB1 et l'arginine déméthylase JMJD6. De façon surprenante, l'étude de l'expression de LKB1 par immunohistochimie dans une cohorte de tumeurs mammaires a montré une dualité fonctionnelle selon sa localisation subcellulaire. De plus, nous avons démontré que JMJD6 s'associe à ERa méthylé lorsque le récepteur est complexé à Src et Pl3K, et permet ainsi la déméthylation de ERa et la dissociation du complexe mERa/Src/Pl3K. Ce travail a ainsi pu mettre en évidence que les différents acteurs de cette signalisation peuvent constituer des éléments clés au diagnostique mais également lors de la décision thérapeutique, puisque qu'il existe des drogues peuvant cibler cette voie de signalisation / Estrogen receptor a {ERa}, belonging to the superfamily of hormone nuclear receptors, regulates many physiological processes, notably the growth and survival of breast tumor cells, acting as a ligand-dependent transcription factor. Besides to the well described transcriptional effects, estrogen also mediate extranuclear events called non genomic signaling via its receptor. /n fact, team shows that ERa is methylated and that this event is a prerequisite for the recrutement of Src and P/3K and the activation of Akt which orchestrate cell proliferation and survival. During my PhD, / demonstrated that the non genomic signaling complex mERa/Src/P/3K exists in vivo and is operative. /n addition, the complex is found to be an independent prognostic factor for disease free survival. This is an emergent concept that estrogen non genomic pathway is operative in vivo and can constitute a new therapeutic targets. The search for new partners of the complex has allowed us to identify the tumor suppressor LKB1 and arginine demethylase JMJD6. Expression of LKB1 in immunohistochemistry revealed dual properties based on its subcellular localization. When LKB1 is complexed with mERa/Src/P/3K it may acquire oncogenic properties. /n addition, JMJD6 interacts with methylated ERa when the receptor is associated with Src and P/3K, and allows the demethylation of ERa and the dissociation of the complex mERa/Src/P/3K. This work showed that estrogenic non genomic players can constitute new therapeutic targets in Breast tumors

L'obésité et ses complications métaboliques comme le syndrome métabolique (SM) deviennent des épidémies mondiales qui partagent une composante commune qu’est l'inflammation chronique. Les acteurs principaux de cette inflammation sont les lymphocytes T. L’utilisation des polyphénols capables de moduler les fonctions de lymphocytes T pour lutter contre les maladies métaboliques inflammatoires fait l’objet de nombreuses investigations. Dans cette étude, nous avons analysé l'effet de la delphinidine, un anthocyane connu pour préserver l'intégrité de l'endothélium par un mécanisme dépendant au récepteur aux oestrogènes alpha (ERα), sur la prolifération, l’apoptose et la différentiation de lymphocytes T isolés à partir de sujets sains et de patients atteints de SM. La delphinidine diminue la prolifération et l’apoptose de lymphocytes T isolés à partir de sujets sains et stimulés par différents agents mitogènes et proapoptotiques, respectivement. Par ailleurs, la delphinidine inhibe la différenciation des lymphocytes vers des profilsTh1, Th17 et Treg, sans affecter le profil Th2. Enfin, la delphinidine inhibe la prolifération, l’apoptose et la différenciation des lymphocytes T isolés à partir de patients présentant des risques cardiovasculaires associés au SM. Les mécanismes moléculaires mis en jeu ont été identifiés, avec l’implication d'ERα, d'ERK1/2, de NFAT et d'HDAC en relation avec le signal calcique. Nos résultats suggèrent que la delphinidine, en agissant sur ERα, via des cibles cellulaires multiples, pourrait représenter une nouvelle approche pour traiter les troubles métaboliques inflammatoires chez les patients présentant des facteurs de risque cardiovasculaire. / Obesity and its metabolic complications like metabolic syndrome (MetS) are becoming worldwide epidemics which share a common component of chronic low-grade inflammation. T lymphocytes play a central role in the triggering of this inflammatory process. Modulation of T lymphocytes functions by using polyphenols, as a possible approach to alleviate chronic inflammatory metabolic diseases has become the subject of many scientific investigations. Here, we analyzed the effect of delphinidin, an anthocyanin known to possess vasculoprotection properties, via an estrogen receptor alpha (ERα)-dependent mechanism, on proliferation, apoptosis and differentiation of T lymphocytes from healthy subjects and MetS patients. We found that delphinidin decreased proliferation and apoptosis of T lymphocytes from healthy subjects stimulated by different mitogen and apoptotic agents, respectively. Further, delphinidin suppressed the differentiation of hese cells toward Th1, Th17 and Treg without affecting Th2 subsets. Interestingly, delphinidin inhibited proliferation, apoptosis and differentiation of T cells taken from patients with cardiovascular risks associated with MetS. We also identified the molecular mechanism involved, with the implication of ERα, ERK1/2, NFAT and HDAC pathways in relation with calcium signaling. Together, we propose that delphinidin by acting on ERα, via multiple cellular targets, may represent a new approach in the treatment of chronic inflammatory metabolic disorders caused by excessive T lymphocyte responses, in with cardiovascular risk factors.

Die Behandlung menopausaler und postmenopausaler Beschwerden ist mit einem erhöhten Risiko verbunden, an Mamma- und Endometriumskarzinomen zu erkranken. Darüber hinaus zeigen epidemiologische Studien, dass in ostasiatischen Ländern postmenopausale Beschwerden, osteoporotische Frakturen und Herz-Kreislauferkrankungen seltener auftreten als in westlichen Ländern. Vor diesem Hintergrund war es Ziel der Untersuchungen dieser Arbeit, in dem Tiermodell der ovarektomierten Ratte die mögliche Bedeutung von estrogenrezeptorsubtypspezifischen Einflüssen für hormonell bedingte Erkrankungen und Beschwerden zu untersuchen. Hierbei sollten gewebespezifische Wirkungen estrogenrezeptorsubtypspezifischer Liganden untersucht und explizit die Bedeutung der Estrogenrezeptorsubtypen ERα und ERβ bei der Gewebehomöostase in Knochen und Darm analysiert werden. Darüber hinaus sollten vor dem Hintergrund estrogenrezeptorsubtypspezifischer Wirkungsweisen gewebespezifische Einflüsse des Phytoestrogens Genistein (Gen) näher charakterisiert werden. Es konnte gezeigt werden, dass nach einer subkutanen Applikation der knochenprotektive Einfluss von Gen mit dem von Estradiol (E2) vergleichbar ist, durch die Kombination mit Bewegung verstärkt wird und über den ERα vermittelt zu sein scheint. Es zeigte sich außerdem, dass der stimulierende Einfluss von E2 auf den motorischen Antrieb ERα-vermittelt ist und ERβ-spezifische Liganden ebenso wie Gen diesen Effekt antagonisieren. Des Weiteren wurde deutlich, dass E2, nicht aber Gen über den ERα Einfluss auf die Körperfettverteilung nimmt. Mit einer phytoestrogenreichen Diät konnten in adulten Ratten physiologisch relevante Gen/Dai-Plasmaspiegel erreicht werden, allerdings blieben hierbei die nach einer subkutanen Applikation beobachteten knochenprotektiven Effekte dieser Phytoestrogene aus. Bei der näheren Betrachtung der Gewebehomöostase im Dünndarm zeigte sich, dass über den im Darm verstärkt exprimierten ERβ antiproliferative und proapoptotische Effekte vermittelt werden und Gen in diesem Gewebe wie ein ERβ-spezifischer Agonist wirkt. Bezogen auf eine hormonell bedingte Osteoporose, wie sie bei einem Großteil postmenopausaler Frauen auftritt, scheint das Phytoestrogen Genistein eine mögliche Alternative zur Hormonersatztherapie darzustellen. Außerdem zeigt sich, dass Genistein gewebe- und estrogenrezeptorsubtypspezifische antagonistische und agonistische Einflüsse hat und somit die Charakterisierung als „Phyto-SERMs“ (pflanzlicher selektiver Estrogenrezeptormodulator) zutreffend ist. Sollten sich in weiterführenden Studien die beobachteten Effekte im Dünndarm auch für die Gewebehomöostase im Kolon beschreiben lassen, können vor diesem Hintergrund Genistein und ERβ-spezifische Liganden für die Darmkrebsprävention diskutiert werden…

Estrogens greatly affect the activity and connectivity of serotonergic neural cell populations, which extend from clusters of nuclei in the brainstem, termed the raphe nuclei, where estrogen receptor β is the most abundantly expressed estrogen receptor subtype. Estrogenic effects on the raphe nuclei are primarily important for influencing various neuropsychological behaviors, including depression, mood swings and anxiety behaviors. Because of this connection, phases of intense hormone fluctuations for instance during menopause are often associated with several mood disturbances that often reduce the quality of life of menopausal women. Accordingly, long-term use of hormone replacement therapy appeared to be the method of choice for many menopausal women to help alleviate vasomotor symptoms, which may include neuropsychological changes such as depression. However, given the limitations and number of serious health risks attributed to hormone replacement therapy, natural compounds such as phytoestrogens are receiving widespread awareness due to their occurrence in medicinal plant extracts and a wide variety of food items including dietary supplements with respective health claims. Flavonoids, particularly the isoflavones and the naringenin-type flavanones, belong to a group of polyphenolic plant-derived secondary metabolites known to possess estrogen-like bioactivities. Nevertheless, little is known about their transactivational activity and their potential to regulate endogenous gene expression of estrogen responsive genes in the raphe nuclei due to the lack of suitable cellular models expressing sufficient amounts of functional estrogen receptor β. Hence, a raphe nuclei-derived cell line that expresses a functional estrogen receptor β was sought as a model to investigate effects of flavonoids in vitro. In this regard, RN46A-B14 cells derived from embryonic day 13 rat medullary raphe nuclei were primarily used in this study as the main cellular model. Nonetheless, expression of endogenous estrogen receptor β in these cells was not sufficient to pursue downstream investigations of estrogen-dependent activities. To overcome this deficit, a rat raphe nuclei-derived in vitro model that overexpresses a functional estrogen receptor β was initially established (herein termed RNDA cells) by stably transducing its parent cell line, RN46A-B14 cells, with a suitable lentiviral expression vector encoding a human estrogen receptor β gene. The stable expression and the functional characterization of the transgenic receptor was confirmed by Western blot analysis and luciferase reporter gene assays, respectively. The same reporter gene assay was used to scrutinize the transactivational activity of the flavonoids in RNDA cells. Key results revealed that Genistein, Daidzein, Equol, Naringenin and 8-Prenylnaringenin demonstrated high transactivational activity in a concentration-dependent manner by stimulating luciferase expression from an estrogen responsive element-regulated reporter gene construct transiently transfected in RNDA cells. Low transactivational activity was observed in RNDA cells in response to increasing concentrations of 7-(O-prenyl)naringenin-4'-acetate. However, no transactivational activity was noticed in response to 6-(1,1-Dimethylallyl)naringenin in the studied cell model. All effects elicited by the flavonoids were antagonized by the pure estrogen receptor antagonist, Fulvestrant, indicating that all substances act by binding to and activating the transgenic ERβ. Additional effects were observed in RNDA cells in response to a co-treatment of 1 µM of either Genistein or Daidzein, but not Equol, with 10 nM 17β-Estradiol. Slight antagonistic effects were observed in the same studied cell line when either 8-Prenylnaringenin or 7-(O-prenyl)naringenin-4'-acetate, but not Naringenin or 6-(1,1-Dimethylallyl)naringenin, were co-added with 17β-Estradiol. Results from the reporter gene assays were validated on the basis of regulation of mRNA expression of estrogen responsive genes following the global assessment of 17β-Estradiol-induced gene expression in this cell line using a DNA microarray technique. Out of 212 estrogen-regulated genes with at least two-fold change of expression, six were selected according to specific features of estrogenic regulation of expression. The expression of the six selected 17β-Estradiol-regulated genes was validated using quantitative real-time PCR analysis. The regulation of mRNA expression of the selected genes in response to the tested flavonoids was then investigated in RNDA cells. Additionally, because RNDA cells encode a temperature-sensitive mutant of the Simian Virus 40 large T-antigen, their neuronal differentiation is constitutive upon shifting them from conditions promoting proliferation (permissive temperature) to differentiation (non permissive temperature). Hence, the regulation of mRNA expression of the selected genes in response to the tested flavonoids was additionally investigated as RNDA cells differentiate. In RNDA cells grown under proliferative conditions, 17β-Estradiol up-regulated mRNA expression of camello-like 5, sex determining region Y-box 18 and keratin type I cytoskeletal 19. Similar effects were observed in response to 8-Prenylnaringenin, Genistein, Daidzein and Equol. In addition, 17β-Estradiol down-regulated mRNA expression of neurofilament medium polypeptide and zinc finger DHHC-type containing 2. Similar effects were observed in response to 8-Prenylnaringenin, Naringenin, Genistein, Daidzein and Equol. Yet, no effect was observed on the regulation of mRNA expression of solute carrier family 6 member 4 in response to 17β-Estradiol or the flavonoids in RNDA cells grown under proliferative conditions. When RNDA cells were shifted to conditions promoting differentiation, changes in cell morphology, in mRNA expression levels and in responsiveness towards 17β-Estradiol or the flavonoids were observed. These expression studies additionally highlighted some of the genes as indicator genes for RNDA cellular differentiation. The newly established RNDA cell line should prove useful to elucidate basic physiological properties of estrogen receptor β in the raphe nuclei. The present study should serve as the basis to help shed light on molecular and cellular mechanisms following the action of phytoestrogens, endocrine disruptors or other exogenous estrogen receptor ligands in neural cell populations, particularly the raphe nuclei, for further applications within the brain.

Carvacrol is a phenol monoterpene found in aromatic plants specially in Lamiaceae family, which has been evaluated in an experimental model of breast cancer. However, any proposed mechanism based on its antitumor effect has not been reported. In our previous study, carvacrol showed a protective effect on 7,12-dimethylbenz[α]anthracene- (DMBA-) induced breast cancer in female rats. The main objective in this research was to evaluate by using in silico study the carvacrol on HER2, PI3Kα, mTOR, hERα, PR, and EGFR receptors involved in breast cancer progression by docking analysis, molecular dynamic, and drug-likeness evaluation. A multilevel computational study to evaluate the antitumor potential of carvacrol focusing on the main targets involved in the breast cancer was carried out. The in silico study starts with protein-ligand docking of carvacrol followed by ligand pathway calculations, molecular dynamic simulations, and molecular mechanics energies combined with the Poisson–Boltzmann (MM/PBSA) calculation of the free energy of binding for carvacrol. As result, the in silico study led to the identification of carvacrol with strong binding affinity on mTOR receptor. Additionally, in silico drug-likeness index for carvacrol showed a good predicted therapeutic profile of druggability. Our findings suggest that mTOR signaling pathway could be responsible for its preventive effect in the breast cancer. / Revisión por pares

No description available.