title: Down-regulation of microRNA-184 contributes to the development of cyanotic congenital heart diseases.
abstract: OBJECTIVE
  We aimed to investigate the roles of miR-184 in adaptation of hypoxic cardiomyocytes, as well as to elucidate the possible mechanisms of miR-184 in the development of cyanotic congenital heart diseases (CHD).
  
  
  METHODS
  We conducted quantitative real-time polymerase chain reaction (qRT-PCR) to determine the expression of miR-184 in patients with cyanotic cardiac defects. The embryonic rat ventricular myocardial H9c2 cells were transfected with miR-184 inhibitor and negative scramble RNA. Mock group was untreated by anything. We then used MTT assay and in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) to determine whether inhibition of miR-184 in vitro affect cell proliferation and apoptosis under hypoxic conditions. Besides, the expression levels of caspase-3 and caspase-9 in hypoxic H9c2 cells were determined by western blot.
  
  
  RESULTS
  MiR-184 was significantly down-regulated in CHD patients with cyanotic cardiac defects. In addition, miR-184 was successfully inhibited in hypoxic H9c2 cells. Moreover, inhibition of miR-184 markedly decreased cell viability and obviously induced apoptosis under hypoxic conditions in vitro. Besides, the expression levels of caspase-3 and caspase-9 in hypoxic H9c2 were significantly increased after miR-184 inhibition.
  
  
  CONCLUSIONS
  Our findings indicate that inhibition of microRNA-184 may contribute to the development of cyanotic CHD via decreasing proliferation and inducing apoptosis of cardiomyocytes. Moreover, miR-184 inhibition may promote hypoxia-induced apoptosis via activation of caspase-3 and caspase-9. Congenital down-regulation of miR-184 may be a mechanism leading to CHD development.
authors.affiliation: Department of Cardiovascular Surgery, The First Hospital of Hebei Medical University Shijiazhuang 050030, P. R. China.
authors.forename: Jiancheng
authors.initials: J
authors.lastname: Huang
authors.affiliation: Department of Cardiovascular Surgery, The First Hospital of Hebei Medical University Shijiazhuang 050030, P. R. China.
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Cardiovascular Surgery, The First Hospital of Hebei Medical University Shijiazhuang 050030, P. R. China.
authors.forename: Hongying
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Cardiovascular Surgery, The First Hospital of Hebei Medical University Shijiazhuang 050030, P. R. China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Su
authors.affiliation: Department of Cardiovascular Surgery, The First Hospital of Hebei Medical University Shijiazhuang 050030, P. R. China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Cardiovascular Surgery, The First Hospital of Hebei Medical University Shijiazhuang 050030, P. R. China.
authors.forename: Huijun
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis; D016022:Case-Control Studies; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D015687:Cell Hypoxia; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D003490:Cyanosis / Q000209:etiology*; D015536:Down-Regulation; D004789:Enzyme Activation; D018507:Gene Expression Regulation, Developmental; D006330:Heart Defects, Congenital / Q000150:complications / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D020287:In Situ Nick-End Labeling; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection
chemical_list: C534424:MIRN184 microRNA, human; C000602309:MIRN184 microRNA, rat; D035683:MicroRNAs; C505410:Casp3 protein, rat; C506435:Casp9 protein, rat; D053148:Caspase 3; D053453:Caspase 9
keywords: Congenital heart diseases; caspase-3; caspase-9; chronic hypoxia; microRNA-184
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