Expression and transcriptional regulation of ABC transporters and cytochromes P450 in hCMEC/D3 human cerebral microvascular endothelial cells.
We investigated the expression of genes encoding ABC transporters, cytochromes P450 (CYPs) and some transcription factors in the hCMEC/D3 immortalized human cerebral microvascular endothelial cell line, a promising in vitro model of the human BBB, and we compared these expressions to a non-brain endothelial cell line (HUVEC) and freshly human brain microvessels. qRT-PCR showed that the MDR1, BCRP, MRP1, MRP3, MRP4 and MRP5 genes were expressed and that the main CYP gene was CYP2U1 in hCMEC/D3. The pattern of ABC and CYPs gene expression in hCMEC/D3 differed from HUVEC which did not express MDR1. Moreover, expression of P-gp and BCRP was lower in hCMEC/D3 than in human brain microvessels but remain functional as shown by rhodamine 123 efflux assay. The gene encoding the aryl hydrocarbon receptor (AhR), a transcription factor that regulates the expression of some ABC and CYPs was highly expressed in hCMEC/D3 and HUVEC, while the pregnane-X-receptor (PXR) and the constitutive androstane receptor (CAR) were barely detected. We investigated the function of the AhR-mediated regulatory pathway in hCMEC/D3 by treating them with the AhR agonist TCDD. The expressions of two AhR-target genes, CYP1A1 and CYP1B1, were increased 26-fold and 28-fold. But the expressions of ABC transporter genes were not significantly altered. We have thus determined the pattern of expression of the genes encoding ABC transporters, CYPs and three transcription factors in hCMEC/D3 and shown that the AhR pathway might afford an original functional transport and metabolic pattern in cerebral endothelial cells that is different from other peripheral endothelial cells.
Dauchy
Sandrine
S
Neuropsychopharmacologie des addictions (CNRS UMR 7157), Université Paris Descartes, Faculté de Pharmacie, Paris, France.
Miller
Florence
F
Couraud
Pierre-Olivier
PO
Weaver
Richard J
RJ
Weksler
Babette
B
Romero
Ignacio-Andres
IA
Scherrmann
Jean-Michel
JM
De Waziers
Isabelle
I
Declèves
Xavier
X
D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D001921:Brain / Q000098:blood supply*; D002461:Cell Line, Transformed; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D017931:DNA Primers; D013759:Dronabinol / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D000072317:Polychlorinated Dibenzodioxins / Q000494:pharmacology; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012293:Rifampin / Q000494:pharmacology; D014158:Transcription, Genetic
D018528:ATP-Binding Cassette Transporters; D017931:DNA Primers; D000072317:Polychlorinated Dibenzodioxins; D012333:RNA, Messenger; D013759:Dronabinol; D003577:Cytochrome P-450 Enzyme System; D012293:Rifampin