title: Genetic variants may play an important role in mRNA-miRNA interaction: evidence for haplotype-dependent downregulation of ABCC2 (MRP2) by miRNA-379.
abstract: BACKGROUND
  The functional influence of single-nucleotide polymorphisms (SNPs) of the ATP-binding cassette (ABC) transporter ABCC2 (MRP2) has been characterized in numerous studies. The aim of this study was to address the question of whether distinct ABCC2 haplotypes, which differ in their mRNA secondary structures, show an influence on the degree of mRNA and protein downregulation through miRNA interaction.
  
  
  METHODS
  A model using human peripheral blood monocytic cells (PBMCs) isolated from healthy Caucasian volunteers, with three defined ABCC2 haplotypes comprising the 5'-UTR SNP -24C>T, the 1249G>A SNP (V417I), and the silent 3972C>T SNP, was outlined. Cells were transiently transfected with miRNA-379, already known to target ABCC2 in HepG2 cells.
  
  
  RESULTS
  ABCC2 was downregulated through miR-379 in a haplotype-dependent manner: the wild-type CGC/CGC was modestly affected (mRNA: -12.7±4.2%, protein: -9.9±0.1%), whereas variant haplotypes were more strongly suppressed: CGT/CGT (mRNA: -36.7±2.4%, protein: -21.6±0.4%) and TGT/TGT (mRNA: -55.7±1.2%, protein: -46.3±4.0%). In addition, glutathione-methylfluorescein efflux was significantly reduced in miR-379-transfected peripheral blood monocytic cells corresponding to ABCC2 protein expression.
  
  
  CONCLUSIONS
  This observation may suggest a differential suppression of ABCC2 by miR-379 caused by haplotype-dependent differences in mRNA secondary structures, resulting in changes in mRNA target accessibility or mRNA stability.
authors.affiliation: Institute for Experimental and Clinical Pharmacology, University Hospital Schleswig-Holstein, Kiel, Germany.
authors.forename: Anneke N
authors.initials: AN
authors.lastname: Werk
authors.forename: Henrike
authors.initials: H
authors.lastname: Bruckmueller
authors.forename: Sierk
authors.initials: S
authors.lastname: Haenisch
authors.forename: Ingolf
authors.initials: I
authors.lastname: Cascorbi
mesh_terms: D000328:Adult; D005260:Female; D005786:Gene Expression Regulation; D006239:Haplotypes; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis* / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D020871:RNA Stability / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*
chemical_list: D035683:MicroRNAs; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D012333:RNA, Messenger; C000715087:ABCC2 protein, human