title: MiR-466b-1-3p regulates P-glycoprotein expression in rat cerebral microvascular endothelial cells.
abstract: Epilepsy is one of the most common neurological disorders, and approximately one-third of epilepsy cases are resistant to treatment with anti-epileptic drug (AED). P-glycoprotein (P-gp) is a multi-drug transporter that is thought to play a pivotal role in multiple drug resistance (MDR) in epilepsy. The regulatory mechanism of P-gp remains largely unknown; however, recent studies have demonstrated that microRNAs (miRNAs) may regulate the chemo-resistance mediated by P-gp. This study investigated the effect of specific miRNAs that regulate P-gp expression in rat cerebral microvascular endothelial cells (RCMECs). Primary cultures of RCMECs were treated with phenobarbital (PB) at various concentrations to induce P-gp overexpression. MiRNA microarrays were used to investigate the expression profiles of miRNAs in the resistant RCMECs induced by PB and corresponding non-resistant cells. Our data demonstrated decreased miR-466b-1-3p expression in the resistant cells compared with the non-resistant cells. Moreover, the recombinant RNA of 466b-1-3p (mimic) and the artificial antisense RNA of miR-466b-1-3p (inhibitor) were constructed and transfected into resistant RCMECs. The expression and function of P-gp were measured by Western blotting, quantitative real-time polymerase chain reaction (qRT-PCR) and flow cytometry using rhodamine efflux. The mRNA and protein levels of P-gp increased as the concentration of PB increased, whereas miR-466b-1-3p levels decreased with increasing PB concentrations (P<0.05). The miR-466b-1-3p mimic down-regulated P-gp expression, whereas the miR-466b-1-3p inhibitor up-regulated P-gp expression (P<0.05). These findings demonstrate that miR-466b-1-3p may regulate PB-induced P-gp expression in RCMECs.
authors.affiliation: Department of Neurology, Jinshan Hospital, Fudan University, Shanghai, China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Yang
authors.affiliation: Department of Center Laboratory, Jinshan Hospital, Fudan University, Shanghai, China; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China.
authors.forename: Weimin
authors.initials: W
authors.lastname: Ren
authors.affiliation: Department of Neurology, Jinshan Hospital, Fudan University, Shanghai, China.
authors.forename: Yiye
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Department of Neurology, Jinshan Hospital, Fudan University, Shanghai, China. Electronic address: yinghuichen@fudan.edu.cn.
authors.forename: Yinghui
authors.initials: Y
authors.lastname: Chen
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics / Q000378:metabolism*; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D000927:Anticonvulsants / Q000494:pharmacology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000098:blood supply*; D004351:Drug Resistance; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000378:metabolism*; D055806:Microvessels / Q000187:drug effects / Q000378:metabolism; D010634:Phenobarbital / Q000494:pharmacology; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000378:metabolism; D017208:Rats, Wistar; D020112:Rhodamine 123 / Q000378:metabolism
chemical_list: D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000927:Anticonvulsants; D005456:Fluorescent Dyes; C000601998:MIRN466 microRNA, rat; D035683:MicroRNAs; D012333:RNA, Messenger; D020112:Rhodamine 123; C095122:multidrug resistance protein 3; D010634:Phenobarbital
keywords: Epilepsy; Multiple drug resistance; P-glycoprotein; Phenobarbital; Rat cerebral microvascular endothelial cell; miR-466b-1-3p