Angiotensin II-Induced Long Non-Coding RNA Alivec Regulates Chondrogenesis in Vascular Smooth Muscle Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animal Studies
2.2. Cell Culture and Treatment
2.3. Treatment with Inhibitors of the AT1R and Signal Transduction Pathways
2.4. RNA Isolation and RT-qPCR
2.5. Cellular Fractionation
2.6. RNA Fluorescence In Situ Hybridization (RNA-FISH)
2.7. 5′ and 3′ Rapid Amplification of cDNA Ends (RACE)
2.8. In Vitro Transcription and Translation
2.9. Transient Transfection of RVSMCs with Plasmids, GapmeRs and siRNAs
2.10. Affymetrix Gene Array Analyses
2.11. Identification of Rat lncRNA Alivec
2.12. Alcian Blue Staining to Determine Chondrogenic Phenotype
2.13. AngII-Infused Rat Model of Hypertension and Vasculopathy
2.14. Tissue Staining and Immunohistochemistry
2.15. Alivec RNA-Pulldown and Mass Spectrometry
2.16. UV-RNA Immunoprecipitation (RIP) Assay
2.17. Data Deposition
2.18. Statistical Analysis
3. Results
3.1. Alivec Is an AngII-Induced lncRNA Adjacent to Chondrogenic Gene Acan in RVSMCs
3.2. AngII-induces Alivec and Acan Expression via Activation of AT1R and Src Kinase
3.3. Alivec Knockdown Attenuates AngII-Induced Upregulation of Genes Associated with Chondrogenesis
3.4. Alivec Mediates a Chondrogenic/Osteogenic Phenotype in RVSMCs
3.5. Transcription Factor Sox9 Controls Alivec Expression in RVSMCs
3.6. Alivec RNA Interacts with hnRNPA2B1 as well as with Tropomyosin alpha-3 Chain, a Protein with Putative Association with the Contractile Phenotype of RVSMCs
3.7. AngII Treatment Increases Aortic Expression of Alivec in Rats
3.8. The Human ALIVEC Locus Contains ACAN Regulatory Elements and a Blood Pressure Quantitative Trait Locus (QTL)
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Samara, V.A.; Das, S.; Reddy, M.A.; Tanwar, V.S.; Stapleton, K.; Leung, A.; Abdollahi, M.; Ganguly, R.; Lanting, L.; Natarajan, R. Angiotensin II-Induced Long Non-Coding RNA Alivec Regulates Chondrogenesis in Vascular Smooth Muscle Cells. Cells 2021, 10, 2696. https://doi.org/10.3390/cells10102696
Samara VA, Das S, Reddy MA, Tanwar VS, Stapleton K, Leung A, Abdollahi M, Ganguly R, Lanting L, Natarajan R. Angiotensin II-Induced Long Non-Coding RNA Alivec Regulates Chondrogenesis in Vascular Smooth Muscle Cells. Cells. 2021; 10(10):2696. https://doi.org/10.3390/cells10102696
Chicago/Turabian StyleSamara, Vishnu Amaram, Sadhan Das, Marpadga A. Reddy, Vinay Singh Tanwar, Kenneth Stapleton, Amy Leung, Maryam Abdollahi, Rituparna Ganguly, Linda Lanting, and Rama Natarajan. 2021. "Angiotensin II-Induced Long Non-Coding RNA Alivec Regulates Chondrogenesis in Vascular Smooth Muscle Cells" Cells 10, no. 10: 2696. https://doi.org/10.3390/cells10102696