Cyclic Electromagnetic DNA Simulation (CEDS) can enhance the Functions of Anticancer miR-26a-5p and miR-126-5p in RAW 264.7 cells

Abstract

Since cyclic electromagnetic DNA simulation (CEDS) was found to increase the hybridization potential of double-stranded DNA (dsDNA) and affect the functions of plasmid DNA (1), in this study, Dodecagonal CEDS was applied to RAW 264.7 cells to target anticancer miRNAs, miR-26a-5p and miR-126-5p, at 20-25 Gauss for 20 min during cell culture, followed by cytological observation, quantitative polymerase chain reaction (qPCR), and immunoprecipitation-based high-performance liquid chromatography (IP-HPLC). Both of miR-26a-5p*-CEDS and miR-126-5p*-CEDS induced significant decrease in cell number, and strong immunoreaction of c-caspase 3 in the remaining cells compared to oncogenic miR-21-5p*-CEDS and untreated control. In qPCR results, miR-26a-5p*-CEDS and miR-126-5p*-CEDS markedly increased the expression of target primary and mature miRNAs. IP-HPLC analysis showed that both miR-26a-5p*-CEDS and miR-126-5p*-CEDS induced potent anticancer effect on RAW 264.7 cells by suppressing RAS signaling, proliferation-growth-cytodifferentiation signaling axis, and angiogenesis-survival-oncogenesis-chronic inflammation axis compared to the untreated control and the positive control performed with non-specific poly-A 12A*-CEDS. MiR-26a-5p*-CEDS increased miRNA biogenesis and ER stress signaling more than miR-126-5p*-CEDS, while miR-126-5p*-CEDS increased NFkB signaling-immunity-apoptosis signaling axis more than miR-26a-5p*-CEDS. Therefore, it is proposed that the sequential treatment of miR-26a-5p*-CEDS and miR-126-5p*-CEDS may give a synergistic anticancer effect on RAW 264.7 cells. It is postulated that CEDS using a miRNA sequence can stimulate hybridization between miRNA and target mRNA and also increase the expression of target miRNA, and subsequently decrease the expression of miRNA target proteins, resulting in the alteration of protein signaling pathways in cells.