Hypoxia Promotes Vascular Smooth Muscle Cell Proliferation through microRNA-Mediated Suppression of Cyclin-Dependent Kinase Inhibitors
Abstract
:1. Introduction
2. Material and Methods
2.1. Cell Culture and Hypoxia
2.2. Quantitative Reverse Transcriptase-PCR (qRT-PCR)
- p115, 5′-CGCCCACAACGATTTATTT-3′ and 5′-TTCGCTTCATGGTGAGTGTC-3′;
- p16, 5′-ATATGCCTTCCCCCACTACC-3′ and 5′-CCCCTGAGCTTCCCTAGTTC-3′;
- p18, 5′-ACGTCAATGCACAAAATGGA-3′ and 5′-TCAGCTTGAAACTCCAGCAA-3′;
- p19, 5′-TGCAGGTCATGATGTTTGG-3′ and 5′-CAGCAGTGTGACCCTCTTGA-3′;
- p21, 5′-GACACCACTGGAGGGTGACT-3′ and 5′-CCACATGGTCTTCCTCTGCT-3′;
- p27, 5′-GAGTGGCAAGAGGTGGAGAA-3′ and 5′-GCGTGTCCTCAGAGTTAGCC-3′;
- p57, 5′-CAGGAGCCTCTCGCTGAC-3′ and 5′-CTTCTCAGGCGCTGATCTCT-3′.
2.3. miRNA Mimics and Anti-miRNA Oligonucleotides
2.4. Luciferase Reporter Constructs
2.5. Luciferase Assay
2.6. Immunoblotting
2.7. Cell Proliferation Assay
2.8. Immunofluorescence Staining
2.9. Statistical Analysis
3. Results
3.1. Hypoxia Downregulates Expression Levels of Specific CDK Inhibitors
3.2. Identification of miRNAs That Target CDK Inhibitors under Hypoxia
3.3. Hypoxia-Induced miRNAs Regulates the Expression of CDK Inhibitors
3.4. Validation of CDK Inhibitors as Novel Targets of miR-497, miR-1268a, and miR-665
3.5. miR-497, miR-1268a, and miR-665 Regulate VSMC Proliferation
3.6. Modulation of miRNAs Regulates the Hypoxia-Induced Proliferation of VSMCs
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Lee, J.; Kang, H. Hypoxia Promotes Vascular Smooth Muscle Cell Proliferation through microRNA-Mediated Suppression of Cyclin-Dependent Kinase Inhibitors. Cells 2019, 8, 802. https://0-doi-org.brum.beds.ac.uk/10.3390/cells8080802
Lee J, Kang H. Hypoxia Promotes Vascular Smooth Muscle Cell Proliferation through microRNA-Mediated Suppression of Cyclin-Dependent Kinase Inhibitors. Cells. 2019; 8(8):802. https://0-doi-org.brum.beds.ac.uk/10.3390/cells8080802
Chicago/Turabian StyleLee, Jihui, and Hara Kang. 2019. "Hypoxia Promotes Vascular Smooth Muscle Cell Proliferation through microRNA-Mediated Suppression of Cyclin-Dependent Kinase Inhibitors" Cells 8, no. 8: 802. https://0-doi-org.brum.beds.ac.uk/10.3390/cells8080802