Roles of microRNA-29a in expression of Bcl-2 and Mcl-1 in rat cardiomyocytes |
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Authors: | ZHANG Zhen-Hui YOU Xiang-Yu LIU Shao-Jun LIU Lian LIU Shi-Ming |
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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. |
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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. |
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Keywords: | MicroRNA-29a Apoptosis B-cell leukemia/lymphoma-2 Myeloid cell leukemia-1 |
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