Department of Orthopaedics, Tianjin Medical University General Hospital, Tianjin, P. R. China.
Department of Orthopaedics, The Affiliated Hospital of Xuzhou, Medical University, Xuzhou, Jiangsu, P. R. China.
J Cell Biochem. 2018 Jul;119(7):5813-5820. doi: 10.1002/jcb.26768. Epub 2018 Apr 10.
Neural stem cells (NSCs) are self-renewing, pluripotent, and undifferentiated cells which have benefits as candidates for central nervous system (CNS) injury. However, the transplanted NSCs usually maintain their undifferentiated characteristics, or differentiated potentially along the glial lineage after transplantation. MicroRNAs (miRNAs) are small, non-coding RNAs that play key roles in cell differentiation. However, it is unknown whether miR-29a could play a role in the process of NSC's differentiation. Primary NSCs were derived from rat embryonic cortex. Lentiviral vector-mediated miR-29a (LV-miR-29a) and negative control (LV-null) were infected into NSCs. Quantitative real-time PCR was used to detect expression of miR-29a and PTEN. Immunocytochemistry was used to stain neurons, astrocytes, and oligodendrocytes. Dual luciferase assay to study the interaction between miR-29a and PTEN. The current study showed that the expression of miR-29a was upregulated during NSC differentiation, while the expression of PTEN was downregulated during NSC differentiation. After infection with LV-miR-29a, MAP-2-positive neurons significantly increased, and GFAP-positive astrocytes significantly decreased. Furthermore, we demonstrated that PTEN is a molecular target of miR-29a. miR-29a promote the neuronal differentiation and decrease the astrocytes differentiation of NSCs via targeting PTEN. This may give insight into a novel mechanism of NSC differentiation and provide a promising therapeutic target.
神经干细胞(NSCs)是自我更新的、多能的和未分化的细胞,它们作为中枢神经系统(CNS)损伤的候选物具有益处。然而,移植的 NSCs 通常保持其未分化的特征,或者在移植后潜在地沿着神经胶质谱系分化。microRNAs(miRNAs)是小的非编码 RNA,在细胞分化中发挥关键作用。然而,miR-29a 是否能在 NSC 分化过程中发挥作用尚不清楚。原代 NSCs 源自大鼠胚胎皮层。慢病毒载体介导的 miR-29a(LV-miR-29a)和阴性对照(LV-null)被感染到 NSCs 中。实时定量 PCR 用于检测 miR-29a 和 PTEN 的表达。免疫细胞化学用于染色神经元、星形胶质细胞和少突胶质细胞。双荧光素酶测定用于研究 miR-29a 和 PTEN 之间的相互作用。本研究表明,miR-29a 在 NSC 分化过程中表达上调,而 PTEN 在 NSC 分化过程中表达下调。感染 LV-miR-29a 后,MAP-2 阳性神经元显著增加,GFAP 阳性星形胶质细胞显著减少。此外,我们证明了 PTEN 是 miR-29a 的分子靶标。miR-29a 通过靶向 PTEN 促进 NSCs 的神经元分化和减少星形胶质细胞分化。这可能为 NSC 分化的新机制提供了启示,并为潜在的治疗靶点提供了依据。