Jurek Benjamin, Denk Lucia, Schäfer Nicole, Salehi Mohammad Saied, Pandamooz Sareh, Haerteis Silke
Institute for Molecular and Cellular Anatomy, University of Regensburg, Regensburg, Germany.
Research Group Neurobiology of Stress Resilience, Max Planck Institute of Psychiatry, Munich, Germany.
Front Cell Neurosci. 2022 Oct 3;16:1000538. doi: 10.3389/fncel.2022.1000538. eCollection 2022.
Oxytocin (OXT) is a neuropeptide that has been associated with neurological diseases like autism, a strong regulating activity on anxiety and stress-related behavior, physiological effects during pregnancy and parenting, and various cellular effects in neoplastic tissue. In this study, we aimed to unravel the underlying mechanism that OXT employs to regulate cell-cell contacts, spheroid formation, and cellular migration in a 3D culture model of human MLS-402 cells. We have generated a labeled OXT receptor (OXTR) overexpressing cell line cultivated in spheroids that were treated with the OXTR agonists OXT, Atosiban, and Thr-Gly-oxytocin (TGOT); with or without a pre-treatment of antisense oligos (Gapmers) that induce exon skipping in the human OXTR gene. This exon skipping leads to the exclusion of exon 4 and therefore a receptor that lost its intracellular G-protein-binding domain. Sensitive digital PCR (dPCR) provided us with the means to differentiate between wild type and truncated OXTR in our cellular model. OXTR truncation differentially activated intracellular signaling cascades related to cell-cell attachment and proliferation like Akt, ERK1/2-RSK1/2, HSP27, STAT1/5, and CREB, as assessed by a Kinase Profiler Assay. Digital and transmission electron microscopy revealed increased tight junction formation and well-organized cellular protrusions into an enlarged extracellular space after OXT treatment, resulting in increased cellular survival. In summary, OXT decreases cellular migration but increases cell-cell contacts and therefore improves nutrient supply. These data reveal a novel cellular effect of OXT that might have implications for degenerating CNS diseases and tumor formation in various tissues.
催产素（OXT）是一种神经肽，与自闭症等神经系统疾病有关，对焦虑和应激相关行为具有强大的调节作用，在怀孕和育儿期间具有生理效应，并且在肿瘤组织中具有多种细胞效应。在本研究中，我们旨在揭示OXT在人MLS-402细胞的三维培养模型中调节细胞间接触、球体形成和细胞迁移的潜在机制。我们构建了一种在球体中培养的过表达标记催产素受体（OXTR）的细胞系，用OXTR激动剂OXT、阿托西班和苏氨酸-甘氨酸-催产素（TGOT）处理；有或没有用反义寡核苷酸（Gapmers）进行预处理，反义寡核苷酸可诱导人OXTR基因外显子跳跃。这种外显子跳跃导致外显子4的缺失，从而产生一种失去细胞内G蛋白结合结构域的受体。灵敏的数字PCR（dPCR）为我们在细胞模型中区分野生型和截短型OXTR提供了方法。通过激酶分析评估，OXTR截短差异激活了与细胞间附着和增殖相关的细胞内信号级联反应，如Akt、ERK1/2-RSK1/2、HSP27、STAT1/5和CREB。数字和透射电子显微镜显示，OXT处理后紧密连接形成增加，细胞突起组织良好，进入扩大的细胞外空间，从而提高细胞存活率。总之，OXT减少细胞迁移，但增加细胞间接触，因此改善营养供应。这些数据揭示了OXT一种新的细胞效应，可能对中枢神经系统退行性疾病和各种组织中的肿瘤形成具有影响。
