title: Suppression of CYP2C9 by microRNA hsa-miR-128-3p in human liver cells and association with hepatocellular carcinoma.
abstract: Published studies have identified genetic variants, somatic mutations, and changes in gene expression profiles that are associated with hepatocellular carcinoma (HCC), particularly involving genes that encode drug metabolizing enzymes (DMEs). CYP2C9, one of the most abundant and important DMEs, is involved in the metabolism of many carcinogens and drugs and is down-regulated in HCC. To investigate the molecular mechanisms that control CYP2C9 expression, we applied integrative approaches including in silico, in vitro, and in vivo analyses to elucidate the role of microRNA hsa-miR-128-3p in the regulation of CYP2C9 expression and translation. RNA electrophoresis mobility shift assays demonstrated a direct interaction between hsa-miR-128-3p and its cognate target, the CYP2C9 transcript. Furthermore, the expression of a luciferase reporter gene containing the 3'-UTR of CYP2C9 and the endogenous expression of CYP2C9 were suppressed by transfection of hsa-miR-128-3p. Importantly, chemically-induced up- or down-regulation of hsa-miR-128-3p correlated inversely with the expression of CYP2C9. Finally, an association analysis revealed that the expression of hsa-miR-128-3p is inversely correlated with the expression of CYP2C9 in HCC tumor tissues. Altogether, the study helped to elucidate the mechanism of CYP2C9 regulation by hsa-miR-128-3p, and the inverse association in HCC.
authors.affiliation: 1] National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA [2] State Key Laboratory of Molecular Oncology and Department of Etiology &Carcinogenesis, Cancer Institute and Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China 100021.
authors.forename: Dianke
authors.initials: D
authors.lastname: Yu
authors.affiliation: National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA.
authors.forename: Bridgett
authors.initials: B
authors.lastname: Green
authors.affiliation: National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA.
authors.forename: April
authors.initials: A
authors.lastname: Marrone
authors.affiliation: Beijing Children's Hospital, Capital Medical University, Beijing, China 100045.
authors.forename: Yongli
authors.initials: Y
authors.lastname: Guo
authors.affiliation: University of Arkansas for Medical Sciences, AR 72205, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Kadlubar
authors.affiliation: State Key Laboratory of Molecular Oncology and Department of Etiology &Carcinogenesis, Cancer Institute and Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China 100021.
authors.forename: Dongxin
authors.initials: D
authors.lastname: Lin
authors.affiliation: National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA.
authors.forename: James
authors.initials: J
authors.lastname: Fuscoe
authors.affiliation: National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA.
authors.forename: Igor
authors.initials: I
authors.lastname: Pogribny
authors.affiliation: National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA.
authors.forename: Baitang
authors.initials: B
authors.lastname: Ning
mesh_terms: D020413:3' Untranslated Regions; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology; D065729:Cytochrome P-450 CYP2C9 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects; D024202:Electrophoretic Mobility Shift Assay; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008113:Liver Neoplasms / Q000235:genetics* / Q000473:pathology; D035683:MicroRNAs / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D016415:Sequence Alignment; D015854:Up-Regulation / Q000187:drug effects; D016047:Zalcitabine / Q000494:pharmacology
chemical_list: D020413:3' Untranslated Regions; C545189:MIRN128 microRNA, human; C542359:MIRN143 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D016047:Zalcitabine; D065729:Cytochrome P-450 CYP2C9