Sharif Samaneh, Ghahremani Mohammad Hossein, Soleimani Masoud
Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
J Biosci. 2017 Dec;42(4):555-563. doi: 10.1007/s12038-017-9714-5.
Neuroblastoma is the most common extracranial solid tumour in children, and differentiation is considered its most appropriate therapy. In this work, we studied effects of miR-124 overexpression on differentiation in M17 cell line as a model of neuroblastoma cancer. Influence of miR-124 overexpression on differentiation in M17 cells was studied. M17 cells were infected with lentivirus that contained miR-124 precursor sequence and followed for 2 weeks to differentiate. Ectopic expression of miR-124 in M17 cells changed the shape of spherical undifferentiated cells to cells with extended neurites that formed neuronal networks. Overexpression of MiR-124 respectively increased the expression level of markers of β-Tubulin III, MAP2, SYN, NF-M and Nestin by 16-, 5-, 4-, 2.3- and 2-folds at the messenger RNA level. MiR-124 overexpression also increased the protein levels of β-Tubulin III and MAP2. Moreover, exogenous expression of miR-124 significantly increased the intracellular calcium in differentiated M17 cells. Since miR-124 is naturally expressed in neuronal cells and is downregulated in neuroblastoma cancer cells, differentiation with this type of microRNA can be a novel treatment for neuroblastoma cancer.
神经母细胞瘤是儿童最常见的颅外实体瘤，分化被认为是其最合适的治疗方法。在这项研究中，我们以M17细胞系作为神经母细胞瘤模型，研究了miR-124过表达对其分化的影响。研究了miR-124过表达对M17细胞分化的影响。用含有miR-124前体序列的慢病毒感染M17细胞，并持续观察2周以使其分化。M17细胞中miR-124的异位表达将球形未分化细胞的形态转变为具有延伸神经突并形成神经网络的细胞。在信使RNA水平上，miR-124的过表达分别使β-微管蛋白III、微管相关蛋白2、突触素、神经丝蛋白M和巢蛋白的标志物表达水平提高了16倍、5倍、4倍、2.3倍和2倍。miR-124过表达还增加了β-微管蛋白III和微管相关蛋白2的蛋白质水平。此外，miR-124的外源表达显著增加了分化的M17细胞内的钙离子浓度。由于miR-124在神经元细胞中天然表达，而在神经母细胞瘤癌细胞中下调，因此用这种类型的微小RNA诱导分化可能是一种治疗神经母细胞瘤的新方法。
