Publication: Biochemical and biophysical research communications
Available evidence suggests the involvement of microRNAs (miRNAs) in the pathological process of several diseases. Nonetheless, molecular mechanism underlying biological effects of miRNAs such as pacemaker exosome-derived miR-127-5p in embryonic-like stem cells (ESCs) differentiation into pacemaker cell is yet to be clarified. Through real time quantitative polymerase chain reaction (qPCR) or western blotting (WB) techniques, levels of miRNAs, miR-127-5p, and NKx2.5 expressions were quantitatively measured. Cellular differentiation (CD) was probed with flow cytometric (FC) and WB techniques. Prediction of miR-127-5p association with NKx2.5 was studied through bioinformatics tools before verification with luciferase assays. Promotion of ESCs differentiation to pacemaker through miR-127-5p was measured with qPCR and WB techniques. Through the same assaying methods, up-regulation of pace-making genes (Shox2, HCN4, Cx45, Tbx3 and Tbx18) expression was observed in Nkx2.5 knockdown group. However, Nkx2.5 expression was down-regulated during differentiation of pacemaker-like cells into ESCs. Furthermore, techniques (such as qPCR, WB, immunofluorescent staining and FC) were employed to demonstrate that overexpressed miR-127-5p could suppress NKx2.5 expression. Through NKx2.5 targeting, ESCs could be differentiated into pacemaker-like cells with miR-127-5p possibly serving as a crucial positive regulator. On the account of our findings, further researches are needed to unearth the possible underlying mechanism and role of exosome-derived miRNAs in cell signaling.