Biomedical Research Institute, Shenzhen Peking University - The Hong Kong University of Science and Technology Medical Center, Shenzhen, China.
Greater Bay Biomedical Innocenter, Shenzhen Bay Laboratory, Shenzhen, China.
J Cell Mol Med. 2021 Jan;25(2):1252-1262. doi: 10.1111/jcmm.16195. Epub 2020 Dec 17.
Pax3 and Pax7 are closely related transcription factors that are widely expressed in the developing nervous system and somites. During the normal development in the central nervous system (CNS), Pax3 and Pax7 are mainly expressed in the dorsal part of the neural tube. Further analysis revealed that Pax3 and Pax7 shared redundant functions in the spinal cord development. However, it is still unknown whether Pax3 and Pax7 play a role in neuronal differentiation. In this study, Pax3 and Pax7 genes were overexpressed in Neuro-2a, the mouse neuroblastoma cell line. CCK-8 and EdU assay results showed that overexpression of Pax3 inhibited cell viability and proliferation of Neuro-2a cells, whereas the overexpression of Pax7 had no significant difference on their cell viability and proliferation. Overexpression of Pax3 not only increased the percentage of cells in the S phase and G0/G1 phase, but also decreased that in the G2 phase. Moreover, the total neurite lengths of Neuro-2a cells were significantly shorter in Pax3 overexpressed group than those in negative control group and showed no significant difference between Pax7 overexpressed group and negative control group. These results suggested that Pax3 not only inhibited the cell viability and proliferation but also affected the cell cycle and the neurite outgrowth of Neuro-2a cells. RNA sequencing analysis showed up-regulated genes in Pax3 overexpressed group were involved in cell cycle machinery, which may reveal the potential mechanism of Neuro-2a cells proliferation.
Pax3 和 Pax7 是密切相关的转录因子，广泛表达于发育中的神经系统和体节中。在中枢神经系统（CNS）的正常发育过程中，Pax3 和 Pax7 主要表达于神经管壁的背侧。进一步的分析显示，Pax3 和 Pax7 在脊髓发育中具有冗余的功能。然而，目前尚不清楚 Pax3 和 Pax7 是否在神经元分化中发挥作用。在这项研究中，过表达 Pax3 和 Pax7 基因于Neuro-2a 细胞系，即小鼠神经母细胞瘤细胞系中。CCK-8 和 EdU 检测结果表明，过表达 Pax3 抑制了Neuro-2a 细胞的活力和增殖，而过表达 Pax7 对其细胞活力和增殖没有显著影响。Pax3 的过表达不仅增加了 S 期和 G0/G1 期细胞的比例，而且降低了 G2 期细胞的比例。此外，过表达 Pax3 的 Neuro-2a 细胞的总神经突长度明显短于阴性对照组，而过表达 Pax7 的组与阴性对照组之间无显著差异。这些结果表明，Pax3 不仅抑制了细胞活力和增殖，而且影响了细胞周期和 Neuro-2a 细胞的神经突生长。RNA 测序分析显示，Pax3 过表达组中上调的基因参与细胞周期机制，这可能揭示了 Neuro-2a 细胞增殖的潜在机制。
