NCAM1 is the Target of miRNA-572: Validation in the Human Oligodendroglial Cell Line.
The neural cell adhesion molecule 1 (NCAM1) is a fundamental protein in cell-cell interaction and in cellular developmental processes, and its dysregulation is involved in a number of diseases including multiple sclerosis. Studies in rats suggest that the modulation of NCAM1 expression is regulated by miRNA-572, but no data are available confirming such interaction in the human system. We analyzed whether this is the case using a human oligodendroglial cell line (MO3.13). MO3.13 cells were transfected with miRNA-572 mimic and inhibitor separately; NCAM1 mRNA and protein expression levels were analyzed at different time points after transfection. Results indicated that NCAM1 expression is increased after transfection with miRNA-572 inhibitor, whereas it is decreased after transfection with the mimic (p < 0.005). The interaction between NCAM1 and miRNA-572 was subsequently confirmed in a Vero cell line that does not express NCAM1, by luciferase assay after transfection with NCAM1. These results confirm that miRNA-572 regulates NCAM1 and for the first time demonstrate that this interaction regulates NCAM1 expression in human cells. Data herein also support the hypothesis that miRNA-572 is involved in diseases associated with NCAM1 deregulation, suggesting its possible use as a biomarker in these diseases.
Don Carlo Gnocchi Foundation IRCCS - ONLUS, Via Capecelatro 66, 20148, Milan, Italy. rmancuso@dongnocchi.it.
Roberta
R
Mancuso
Don Carlo Gnocchi Foundation IRCCS - ONLUS, Via Capecelatro 66, 20148, Milan, Italy.
Simone
S
Agostini
Don Carlo Gnocchi Foundation IRCCS - ONLUS, Via Capecelatro 66, 20148, Milan, Italy.
Ivana
I
Marventano
Don Carlo Gnocchi Foundation IRCCS - ONLUS, Via Capecelatro 66, 20148, Milan, Italy.
Ambra
A
Hernis
Don Carlo Gnocchi Foundation IRCCS - ONLUS, Via Capecelatro 66, 20148, Milan, Italy.
Marina
M
Saresella
Don Carlo Gnocchi Foundation IRCCS - ONLUS, Via Capecelatro 66, 20148, Milan, Italy.|Department of Physiopathology and Transplantation, University of Milan, Milan, Italy.
Mario
M
Clerici
D000818:Animals; D019002:CD56 Antigen / Q000096:biosynthesis* / Q000235:genetics*; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics*; D009836:Oligodendroglia / Q000378:metabolism*; D015203:Reproducibility of Results; D014709:Vero Cells
D019002:CD56 Antigen; C000603872:MIRN-572 microRNA, human; D035683:MicroRNAs; C522918:NCAM1 protein, human
Human oligodendrocytes; Multiple sclerosis; Myelination; Neuronal cell adhesion molecule; Transfection; microRNA