Aryl hydrocarbon receptor (AHR) and its heterodimer aryl hydrocarbon nuclear translocator (ARNT) form a ligand-activated transcription complex that regulates expression of the AHR battery of target genes that includes the most important placental biotransformation enzyme cytochrome CYP1A1. Expression, placental localization, and ontogeny of AHR/Ahr and ARNT/Arnt have not been systematically studied in either human or rat placentas. Moreover, induction of such AHR target genes as CYP1A1, CYP1A2, CYP1B1, UGT1A1, and breast cancer resistance protein (BCRP), as well as of AHR, ARNT, and aryl hydrocarbon receptor repressor (AHRR) genes, after exposure to AHR ligands have not been studied in human placental trophoblast cultures. In this article, we show that only CYP1A1 messenger RNA (mRNA), but not CYP1A2, CYP1B1, UGT1A1, BCRP, AHR, ARNT, and AHRR mRNAs, is significantly induced in human term placental trophoblast cultures after exposure to prototype AHR ligands/activators 2,3,7,8-tetrachlorodibenzo-p-dioxin, 3-methylcholanthrene, omeprazole, and β-naphthoflavone. We localized AHR/Ahr and ARNT/Arnt in rat placental trophoblasts throughout gestation and in first trimester and term human placental trophoblast, which comprise the crucial component of the maternal-fetal barrier. We demonstrate that rat Ahr and Cyp1a1 reached highest expression during gestation days 15 and 18, which might indicate different response to Ahr ligands in placental Cyp1a1 induction during rat gestation. We also propose the JEG3 choriocarcinoma cell line as a cellular model for human trophoblast induction studies through AHR. In conclusion, we describe expression and ontogeny of AHR/Ahr and ARNT/Arnt and systematically characterize induction of major AHR target genes in human placental trophoblast forming the placental maternal-fetal morphological and metabolic barrier.

• MeSH
• aktivace transkripce účinky léků   fyziologie   MeSH
• aromatické hydroxylasy genetika   metabolismus   MeSH
• choriokarcinom farmakoterapie   genetika   metabolismus   MeSH
• dospělí MeSH
• gestační stáří MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• nádorové buněčné linie MeSH
• potkani Wistar MeSH
• receptory aromatických uhlovodíků - jaderný translokátor genetika   metabolismus   MeSH
• receptory aromatických uhlovodíků genetika   metabolismus   MeSH
• těhotenství MeSH
• trofoblasty účinky léků   metabolismus   MeSH
• vývojová regulace genové exprese MeSH
• xenobiotika toxicita   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aromatické hydroxylasy MeSH
• messenger RNA MeSH
• receptory aromatických uhlovodíků - jaderný translokátor MeSH
• receptory aromatických uhlovodíků MeSH
• xenobiotika MeSH

Vitamin D receptor (VDR) regulates the expression of many genes involved in mineral metabolism, cellular proliferation, differentiation and drug biotransformation. We studied the expression and activity of VDR and its heterodimerization partner retinoid X receptor-alpha (RXRalpha) in choriocarcinoma trophoblast cell lines BeWo and JEG-3, in comparison with human isolated placental cytotrophoblasts and human full term placenta. We found that VDR and RXRalpha are localised in the human term placenta trophoblast and expressed in isolated cytotrophoblasts. However, we found low expression and no transcriptional activity of VDR in used choriocarcinoma cell lines. The inhibitor of DNA methylation, 5-deoxy-3'-azacytidine, and histone deacetylase inhibitor sodium butyrate partially restored the expression of VDR, suggesting an epigenetic suppression of the gene in choriocarcinoma cells. Differentiation of BeWo cells resulted in up-regulation of VDR mRNA. Finally, we observed a non-genomic effect of 1,25(OH)(2)D(3) in the activation of the extracellular signal-regulated kinase (ERK) signalling pathway in JEG-3 cells. In conclusion, our results suggest an epigenetic repression of VDR gene expression and activity in choriocarcinoma cell lines, and a non-genomic effect of 1,25(OH)(2)D(3) in JEG-3 cells.

• MeSH
• aktivace transkripce účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• choriokarcinom genetika   metabolismus   MeSH
• epigeneze genetická účinky léků   MeSH
• estradiol farmakologie   MeSH
• genom lidský genetika   MeSH
• kadheriny genetika   metabolismus   MeSH
• kalcitriol farmakologie   MeSH
• lidé MeSH
• luciferasy metabolismus   MeSH
• messenger RNA genetika   metabolismus   MeSH
• nádorové buněčné linie MeSH
• placenta cytologie   účinky léků   metabolismus   MeSH
• receptory kalcitriolu genetika   metabolismus   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• reportérové geny MeSH
• retinoidní X receptor alfa genetika   metabolismus   MeSH
• separace buněk MeSH
• těhotenství MeSH
• transport proteinů účinky léků   MeSH
• trofoblasty cytologie   účinky léků   metabolismus   MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• estradiol MeSH
• kadheriny MeSH
• kalcitriol MeSH
• luciferasy MeSH
• messenger RNA MeSH
• receptory kalcitriolu MeSH
• retinoidní X receptor alfa MeSH

1. Breast cancer resistance protein (BCRP, ABCG2) is a drug efflux transporter that is believed to affect the drug disposition of several drugs and xenobiotics. In the present study, we evaluated the localization and functional expression of BCRP in the human choriocarcinoma cell line BeWo, an in vitro model of the human trophoblast, and compared it with the expression of P-glycoprotein (MDR1, ABCB1) as the most widely studied placental transporter. In addition, the expression of BCRP at the mRNA level was compared with that of MDR1 in the human term placenta. 2. Western blotting analysis revealed high endogenous expression of BCRP protein in BeWo cells. Using indirect immunofluorescence microscopy, we found that the BCRP transporter appears to be localized predominantly at the apical plasma membrane. Functional studies showed a significant effect of the BCRP inhibitors GF120918 (5 micromol/L) and Ko143 (1 micromol/L) on mitoxantrone accumulation and, thus, confirmed efflux activity of BCRP in BeWo cells. 3. Using absolute mRNA quantification with real-time reverse transcription-polymerase chain reaction, we found high expression of BCRP in BeWo cells, whereas no transcript of MDR1 (P-glycoprotein), the most extensively studied drug transporter, was detected. 4. In the human placenta, BCRP was localized predominantly in the syncytiotrophoblast layer; however, immunopositivity for the BXP-21 antibody was also observed in fetal vessels of the chorionic villi. The number of BCRP transcripts in the human term placenta was found to be more than 10-fold higher compared with the expression of MDR1. 5. In conclusion, we suggest that BeWo cells could serve as a suitable in vitro model to study trans-trophoblast transport of BCRP substrates and that placental BCRP can play an important role in preventing the accumulation of potentially toxic xenobiotics in the trophoblast cells.

• MeSH
• ABC transportér z rodiny G, člen 2 MeSH
• ABC transportéry antagonisté a inhibitory   genetika   metabolismus   MeSH
• akridiny farmakologie   MeSH
• biologický transport účinky léků   MeSH
• buněčná membrána metabolismus   MeSH
• buněčné linie MeSH
• choriokarcinom genetika   metabolismus   patologie   MeSH
• fluorescenční mikroskopie MeSH
• imunohistochemie MeSH
• indoly chemie   farmakologie   MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• mitoxantron farmakokinetika   MeSH
• nádorové buněčné linie MeSH
• nádorové proteiny antagonisté a inhibitory   genetika   metabolismus   MeSH
• P-glykoprotein genetika   metabolismus   MeSH
• placenta metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• regulace genové exprese u nádorů * MeSH
• těhotenství MeSH
• tetrahydroisochinoliny farmakologie   MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ABC transportér z rodiny G, člen 2 MeSH
• ABC transportéry MeSH
• ABCG2 protein, human MeSH Prohlížeč
• akridiny MeSH
• Elacridar MeSH Prohlížeč
• indoly MeSH
• messenger RNA MeSH
• mitoxantron MeSH
• nádorové proteiny MeSH
• P-glykoprotein MeSH
• tetrahydroisochinoliny MeSH
• tryptoquivaline MeSH Prohlížeč