Li Hong, Ding Chenyue, Ding Zhi-Liang, Ling Mingfa, Wang Ting, Wang Wei, Huang Boxian
Center of Reproduction and Genetics, Suzhou Hospital Affiliated to Nanjing Medical University, Suzhou, China.
Department of Neurosurgery, Suzhou Hospital Affiliated to Nanjing Medical University, Suzhou, China.
J Cell Mol Med. 2017 Aug;21(8):1605-1618. doi: 10.1111/jcmm.13090. Epub 2017 Feb 28.
Human embryonic stem cells (hESCs) can self-renew and differentiate into all cell lineages. E2 is known to exhibit positive effects on embryo development. Although the importance of E2 in many physiological processes has been reported, to date few researchers have investigated the effects of E2 on hESCs differentiation. We studied the effects of E2 on dopamine (DA) neuron induction of hESCs and its related signalling pathways using the three-stage protocol. In our study, 0.1 μM E2 were applied to hESCs-derived human embryoid bodies (hEBs) and effects of E2 on neural cells differentiation were investigated. Protein and mRNA level assay indicated that E2 up-regulated the expression of insulin-like growth factors (IGF)-1, ectoderm, neural precursor cells (NPC) and DA neuron markers, respectively. The population of hESC-derived NPCs and DA neurons was increased to 92% and 93% to that of DMSO group, respectively. Furthermore, yield of DA neuron-secreted tyrosine hydroxylase (TH) and dopamine was also increased. E2-caused promotion was relieved in single inhibitor (ICI or JB1) group partly, and E2 effects were repressed more stronger in inhibitors combination (ICI plus JB1) group than in single inhibitor group at hEBs, hNPCs and hDA neurons stages. Owing to oestrogen receptors regulate multiple brain functions, when single or two inhibitors were used to treat neural differentiation stage, we found that oestrogen receptor (ER)β but not ERα is strongly repressed at the hNPCs and hDA neurons stage. These findings, for the first time, demonstrate the molecular cascade and related cell biology events involved in E2-improved hNPC and hDA neuron differentiation through cross-talk between IGF-1 and ERβ in vitro.
人类胚胎干细胞（hESCs）能够自我更新并分化为所有细胞谱系。已知E2对胚胎发育具有积极作用。尽管已有报道称E2在许多生理过程中具有重要性，但迄今为止，很少有研究人员探究E2对hESCs分化的影响。我们使用三阶段方案研究了E2对hESCs向多巴胺（DA）神经元诱导分化及其相关信号通路的影响。在我们的研究中，将0.1μM的E2应用于hESCs来源的人类胚状体（hEBs），并研究了E2对神经细胞分化的影响。蛋白质和mRNA水平检测表明，E2分别上调了胰岛素样生长因子（IGF）-1、外胚层、神经前体细胞（NPC）和DA神经元标志物的表达。hESC来源的NPC和DA神经元的数量分别增加至二甲基亚砜（DMSO）组的92%和93%。此外，DA神经元分泌的酪氨酸羟化酶（TH）和多巴胺的产量也有所增加。在单一抑制剂（ICI或JB1）组中，E2引起的促进作用部分得到缓解，并且在hEBs、hNPCs和hDA神经元阶段，与单一抑制剂组相比，抑制剂联合（ICI加JB1）组对E2作用的抑制作用更强。由于雌激素受体调节多种脑功能，当使用单一或两种抑制剂处理神经分化阶段时，我们发现在hNPCs和hDA神经元阶段，雌激素受体（ER）β而非ERα受到强烈抑制。这些发现首次证明了在体外，通过IGF-1与ERβ之间的相互作用，E2促进hNPCs和hDA神经元分化所涉及的分子级联反应和相关细胞生物学事件。
