Don Carlo Gnocchi Foundation IRCCS - ONLUS, Via Capecelatro 66, 20148, Milan, Italy.
Department of Physiopathology and Transplantation, University of Milan, Milan, Italy.
Cell Mol Neurobiol. 2018 Mar;38(2):431-440. doi: 10.1007/s10571-017-0486-0. Epub 2017 Mar 22.
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.
神经细胞黏附分子 1(NCAM1)是细胞间相互作用和细胞发育过程中的基本蛋白,其失调与包括多发性硬化症在内的多种疾病有关。大鼠研究表明,NCAM1 表达的调节受 miRNA-572 的调控,但在人类系统中尚无证实这种相互作用的相关数据。我们使用人少突胶质细胞系(MO3.13)对此进行了分析。MO3.13 细胞分别用 miRNA-572 模拟物和抑制剂转染;转染后不同时间点分析 NCAM1 mRNA 和蛋白表达水平。结果表明,miRNA-572 抑制剂转染后 NCAM1 表达增加,而模拟物转染后 NCAM1 表达减少(p<0.005)。随后通过转染 NCAM1 后在不表达 NCAM1 的 Vero 细胞系中进行荧光素酶测定,证实了 NCAM1 和 miRNA-572 之间的相互作用。这些结果证实了 miRNA-572 调节 NCAM1,并且首次证明这种相互作用调节人类细胞中的 NCAM1 表达。本文中的数据还支持了 miRNA-572 参与与 NCAM1 失调相关疾病的假说,表明其在这些疾病中可能作为生物标志物的潜在用途。