Dynamic changes of nuclear RNA foci in proliferating DM1 cells.
Dynamic changes of nuclear RNA foci in proliferating DM1 cells.
复制标题
增殖DM1细胞核RNA焦点的动态变化。
DOI：
10.1007/s00418-015-1315-5
复制
发表时间：
2015
影响因子：
2.3
通讯作者：
Ashizawa,Tetsuo
中科院分区：
文献类型：
--
作者：
Xia,Guangbin;Ashizawa,Tetsuo
核RNA灶是强直性肌营养不良1型（DM1）的分子标志。然而，没有指定的研究已经调查了它们的形成和增殖细胞中的变化。造血干细胞是人体内重要的细胞库。这些细胞中病灶变化的揭示可能揭示突变对这些特定细胞和组织的影响。在本研究中，我们使用人DM1 iPS细胞来源的神经干细胞（NSCs）作为细胞模型来研究增殖细胞中RNA灶的形成和动态变化。在丝裂霉素C处理后，在增殖条件和非增殖条件下培养源自人DM1 iPS细胞的人DM1 NSC。用荧光原位杂交技术观察细胞周期中病灶的动态变化。我们发现RNA灶在细胞周期中形成和解离。核RNA灶在进入有丝分裂（早期前期）之前在数量和大小上最突出。在有丝分裂过程中，大部分病灶消失。进入间期后，RNA灶再次聚集在细胞核内。用丝裂霉素C处理细胞使其停止分裂后，核内RNA灶的数目明显增加。总之，DM1 NSC核RNA焦点在细胞周期中经历动态变化，有丝分裂是减少核中焦点负载的机制，这可以解释为什么分裂细胞受突变影响较小。以病灶作为监测药物疗效的指标时，应考虑病灶的动态变化。
Nuclear RNA foci are molecular hallmarks of myotonic dystrophy type 1 (DM1). However, no designated study has investigated their formation and changes in proliferating cells. Proliferating cells, as stem cells, consist of an important cellular pool in the human body. The revelation of foci changes in these cells might shed light on the effects of the mutation on these specific cells and tissues. In this study, we used human DM1 iPS-cell-derived neural stem cells (NSCs) as cellular models to investigate the formation and dynamic changes of RNA foci in proliferating cells. Human DM1 NSCs derived from human DM1 iPS cells were cultured under proliferation conditions and nonproliferation conditions following mitomycin C treatment. The dynamic changes of foci during the cell cycle were investigated by fluorescence in situ hybridization. We found RNA foci formed and dissociated during the cell cycle. Nuclear RNA foci were most prominent in number and size just prior to entering mitosis (early prophase). During mitosis, most foci disappeared. After entering interphase, RNA foci accumulated again in the nuclei. After stopping cell dividing by treatment of mitomycin C, the number of nuclear RNA foci increased significantly. In summary, DM1 NSC nuclear RNA foci undergo dynamic changes during cell cycle, and mitosis is a mechanism to decrease foci load in the nuclei, which may explain why dividing cells are less affected by the mutation. The dynamic changes need to be considered when using foci as a marker to monitor the effects of therapeutic drugs.