Screening And Identification Of HERG-related MicroRNA

Background and Objective: Malignant arrhythmia is the major cause of sudden cardiacdeath (Sudden Cardiac Death, SCD). The core mechanism of malignant arrhythmia is stillcompletely unknown that why the clinical prevention and treatment are not effective. Themolecular biological research of Long QT Syndrome (LQTS) has provided a new insight for themechanism of malignant arrhythmia. For example, the one of most significant pathogenesis ofLQTS is the alteration of genetic function of the human ether-a-go-go-related gene (hERG),whichis important for encoding the α-subunit of the rapidly activating delayed rectifier K+current (Ikr).Afew of previous studies have demonstrated that abnormal expression of microRNA (miRNA) hasbeen associated with modulation of genetic function of hERG pathway. In this study, we focusedon screening the hERG-related miRNAs by bioinformatics assay, and then validated the hERG-related miRNAs by application of dual-luciferase reporting gene technology. It provided anexperimental basis for later studies to concern on the regulation of miRNA to hERG gene. Inaddition, it also provided a new idea to assess the mechanism of LQTS. Furthermore, it was usefulfor developing much safer and more specific drugs to against arrhythmic.Methods: We predicted the hERG-related miRNA, and synthesized its mimics throughbioinformatics assay. And then we built up the hERG-related dual-luciferase vectors. According tothe manufacturer’s instruction, dual-luciferase vector, dual-luciferase vector and miRNA negativecontrol plasmids or dual-luciferase vector and miRNA mimics were transfected into hela cells byLipofectamine2000. Finally, detection of the activity of relative luciferase, we validated theexact hERG-related miRNA.Results:(1) Used bioinformatics assay, we predicted six hERG-related miRNA: has-miR-134,has-miR-143,has-miR-103a-1,has-miR-147,has-miR-185,has-miR-3619;(2) When miR-143co-transfected with pmirGLO-143, which was the luciferase expression vector of miR-143, it wasfound that the activity of luciferase of miR-143was decreased almost27.12%, which comparedwith blank control group; It is suggested that miR-143was able to inhibit its correspondingluciferase vector pmirGLO-143to express;(3)While miR-103a-1co-transfected with itscorresponding luciferase expression vector pmirGLO-103a-1, to compared with blank controlgroup, the activity of luciferase of miR-103a-1was reduced about34.71%, it was suggested thatmiR-103a-1had the capacity to inhibit its corresponding luciferase vector pmirGLO-103a-1toexpress;(4) when miR-134co-transfected with its corresponding luciferase vector pmirGLO-134, the measure of the luciferase activity compared with blank control group was decreased about20.58%, it was implied that miR-134inhibited the expression of its corresponding luciferasevector pmirGLO-134;(5) when miR-3619co-transfected with its corresponding luciferase vectorpmirGLO-3619, the luciferase activity compared with blank control group decreased by about23.67%; it was pointed out that miR-3619having inhibition on its corresponding luciferaseexpression vector pmirGLO-3619;Conclusion: Our findings illustrated that miR-143、miR-103a-1、miR-134and miR-3619isthe hERG-related miRNA.