Investigating the interaction of hsa-miR-193b-5p and hsa-circ-cnot11-0001 in multiple sclerosis disease through dual luciferase assay

Abstract

Background: Multiple sclerosis (MS) is a chronic autoimmune disorder characterized by dynamic neurodegeneration, basically affecting people between the ages of 20 and 40. Despite significant progress in MS research, little is understood about the underlying molecular mechanisms, particularly those related to epigenetic regulation. According to recent studies, non-coding RNAs (ncRNAs) are crucial epigenetic regulators that have a big impact on how multiple sclerosis develops and is controlled. Given the increasing interest in ncRNAs as biomarkers and therapeutic targets, it is more crucial than ever to look into their role in MS. Recent transcriptomic research showing that circRNAs and miRNAs express differently in neurological disorders makes this especially pertinent. In order to shed light on their possible regulatory role in MS, this study examines the expression levels of circ-cnot11-0001 and hsa-miR-193b-5p in patients and uses a dual luciferase assay to analyze their direct interaction. Methods: The gene expression profile GSE61741 was used to select key miRNAs and circRNAs in MS. Expressions of hsa-miR-193b-5p and circ-cnot11-0001 were verified by RT-qPCR from blood samples of 30 MS patients and 30 controls. After that, we constructed pBud-EGFP plasmid with hsa-miR-193b-5p and psiCHECK-2 plasmid with circ-cnot11-0001. Their direct interaction was determined by measuring luciferase activity. Result: Expression analysis revealed significant downregulation of hsa-miR-193b-5p and upregulation of hsa-circ-cnot11-0001 in MS patients compared to controls. Also, the results of the luciferase activity determined that the luciferase activity of the hsa-miR-193b-5p-pBud + cnot11-0001-WT group was significantly lower than that of the hsa-miR-193b-5p-pBud + cnot11-0001-MT group. So, circ-cnot11-0001 can direct interaction with hsa-miR-193b-5p. Conclusion: The findings indicate a direct interaction between hsa-miR-193b-5p and circ-cnot11-0001 in MS. Therefore, circ-cnot11-0001 may sponge hsa-miR-193b-5p and dysregulate the expression of genes related to MS. In conclusion, the CNOT11-0001-hsa-miR-193b-5p network may serve as a combined biomarker for further investigation into MS.