MicroRNA-153-3p Inhibits Cell Proliferation and Increases Apoptosis in Human Vascular Smooth Muscle Cells in Peripheral Arterial Disease
JOURNAL OF BIOMATERIALS AND TISSUE ENGINEERING(2020)
摘要
Objective: This study explored the role and molecular mechanism of microRNA-153-3p (miR-153-3p) in human vascular smooth muscle cells in peripheral arterial disease. Methods: Human vascular smooth muscle cells (hVSMCs) were transfected with mimic control, miR-153-3p mimic, inhibitor control or miR-153-3p inhibitor for 48 h, and qRT-PCR was applied to determine the transfection efficiency. Meanwhile, the cell viability, cell migration and apoptosis of the transfected cells were measured via MTT assay, flow cytometry analysis and Transwell assay respectively. The target gene of miR-153-3p was predicted by TargetScan and verified by the dual luciferase reporter gene system. In addition, mimic control, miR-153-3p mimic, miR-153-3p mimic+control-plasmid, or miR-153-3p mimic+BCL2-plasmid was transfected into hVSMCs for 48 h, and the cell viability, migration and apoptosis were determined. Besides, qRT-PCR and Western blot assay were applied to measure the expression of BCL2 at both mRNA and protein levels in hVSMCs after the transfection. Results: Compared with the control group, miR-153-3p mimic improved the expression of miR-153-3p, and miR-153-3p inhibitor down-regulated miR-153-3p in hVSMCs. miR-153-3p mimic could reduce the cell viability and migration, and induce the cell apoptosis of hVSMCs. However, miR-153-3p inhibitor had the opposite effects on the cell viability, migration and apoptosis on hVSMCs. We predicted and verified that BCL2 was a direct target of miR-153-3p. In addition, all the effects of miR-153-3p mimic on hVSMCs were significantly reversed by BCL2-plasmid. Conclusion: We demonstrated that miR-153-3p participated in the development and progression of peripheral arterial disease through regulating the proliferation and migration of vascular smooth muscle cells by targeting BCL2.
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关键词
Peripheral Arterial Disease,Human Vascular Smooth Muscle Cells,MicroRNA-153-3p,BCL2
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