Searching the online miRNA database (www.mirdb.org), they identified the miR-29a-3p binding sequence within the 3’-UTR of the target gene and conducted luciferase assay to verify it. Cells were transfected with miR-29a-3p and ICa,L was determined in those cells.
After validating CACNA1C to be the direct target gene of miR-29a-3p, researchers established the negative regulatory relationship between miR-29a-3p and by studying the relative luciferase activity. They also conducted real-time PCR, miRNA microarray and Western blot analysis to study the mRNA and protein expression level of CACNA1C among different groups of cells treated with scramble control, 30nM miR-29a-3p mimics, and 60nM miR-29a-3p mimics, indicating a negative regulatory relationship between miR-29a-3p and CACNA1C. They next analyzed whether miR-29a-3p transfection in cardiomyocytes produced the effects on the ICa,L induced by electrical remodeling, and found a tonic inhibition of IBa by endogenous miR-29a-3p in atrial myocytes.
Through their work, researchers validated the negative regulation between miR-29a-3p and CACNA1C, and found that miR-29a-3p might be a potential therapeutic target in the treatment of AF.
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Y. Zhao, Y. Yuan, C. Qiu (2016) Underexpression of CACNA1C Caused by Overexpression of microRNA-29a Underlies the Pathogenesis of Atrial Fibrillation Med Sci Monit 22:2175-2181 doi: 10.12659/MSM.896191 [article]