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Roles of microRNA-29a in expression of Bcl-2 and Mcl-1 in rat cardiomyocytes
Authors:ZHANG Zhen-Hui  YOU Xiang-Yu  LIU Shao-Jun  LIU Lian  LIU Shi-Ming
Institution:1.Intensive Care Unit, ;2.Guangzhou Institute of Cardiovascular Diseases, ;3.Department of Cardiology, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China.
Abstract:AIM: To investigate the regulatory mechanisms of microRNA-29a (miR-29a) on the expression of Bcl-2 and Mcl-1 in rat cardiomyocytes (CM cells). METHODS: The CM cells were isolated from the hearts of newborn rats and transfected with miR-29a mimic (100 nmol/L) by Lipofectamine RNAiMAX. The expression of Bcl-2 and Mcl-1 at mRNA and protein levels was detected by real-time fluorescence quantitative PCR and Western blotting. The luciferase assay was performed in HEK293T cells and CM cells, which were co-transfected with plasmid DNA and miRNA using Lipofectamine 2000. RESULTS: Transfection of miR-29a mimics significantly reduced the expression levels of Bcl-2 and Mcl-1 in CM cells as compared with the control cells (P<0.05). In addition, HEK293T cells co-transfected with miR-29a mimic and Bcl-2-3’UTR-WT or Mcl-1-3’UTR-WT plasmid significantly reduced the luciferase activity as compared with control group (P<0.05). While CM cells transfected with miR-29a inhibitor and Bcl-2-3’UTR-WT or Mcl-1-3’UTR-WT plasmid in succession, the luciferase activity was increased inversely (P<0.05). CONCLUSION: miR-29a may regulate apoptosis by targeting the bcl-2 and mcl-1 genes.
Keywords:MicroRNA-29a  Apoptosis  B-cell leukemia/lymphoma-2  Myeloid cell leukemia-1  
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