Shanghai Tenth People's Hospital of Tongji University, Tongji University, Middle Yanchang Rd. 301#, Jingan District, Shanghai, China 200072.
Neural Plast. 2020 Nov 25;2020:8855822. doi: 10.1155/2020/8855822. eCollection 2020.
Microglia activation contributes to Alzheimer's disease (AD) etiology, and microglia migration is a fundamental function during microglia activation. The repressor element-1 silencing transcription factor (REST), a powerful transcriptional factor, was found to play a neuroprotective role in AD. Despite its possible role in disease progression, little is known about whether REST participates in microglia migration. In this study, we aimed to explore the function of REST and its molecular basis during microglia migration under A -treated pathological conditions. When treated by A REST was upregulated through JAK2/STAT3 signal pathway in BV2 cells. And transwell coculture system was used to evaluate cell migration function of microglia-like BV2. Small interfering RNA (siRNA) targeting progranulin (PGRN) were delivered into BV2 cells, and results showed that PGRN functions to promote BV2 migration. REST expression was inhibited by sh-RNA, which induced BV2 cell migration obviously. On the contrary, REST was overexpressed by REST recombinant plasmid transfection, which repressed BV2 cell migration, indicating that REST may act as a repressor of cell migration. To more comprehensively examine the molecular basis, we analyzed the promoter sequence of PGRN and found that it has the potential binding site of REST. Moreover, knocking-down of REST can increase the expression of PGRN, which confirms the inhibiting effect of REST on PGRN expression. Further detection of double luciferase reporter gene also confirmed the inhibition of REST on the activity of PGRN promoter, indicating that REST may be an inhibitory transcription factor of PGRN which governs microglia-like BV2 cell migration. In conclusion, the present study demonstrates that transcription factor REST may act as a repressor of microglia migration through PGRN.
小胶质细胞的激活导致阿尔茨海默病（AD）的发生，而小胶质细胞的迁移是小胶质细胞激活的基本功能。抑制元件-1 沉默转录因子（REST）是一种强大的转录因子，被发现对 AD 具有神经保护作用。尽管它在疾病进展中可能发挥作用，但REST 是否参与小胶质细胞迁移知之甚少。在这项研究中，我们旨在探讨 REST 在 A 处理的病理条件下小胶质细胞迁移中的功能及其分子基础。当用 A 处理时，BV2 细胞中的 JAK2/STAT3 信号通路上调了 REST。并使用 Transwell 共培养系统评估了小胶质细胞样 BV2 的细胞迁移功能。将针对颗粒蛋白前体（PGRN）的小干扰 RNA（siRNA）递送至 BV2 细胞，结果表明 PGRN 可促进 BV2 迁移。通过 sh-RNA 抑制 REST 表达，明显诱导 BV2 细胞迁移。相反，通过 REST 重组质粒转染过表达 REST，抑制 BV2 细胞迁移，表明 REST 可能作为细胞迁移的抑制剂。为了更全面地检查分子基础，我们分析了 PGRN 的启动子序列，发现它具有 REST 的潜在结合位点。此外，敲低 REST 可以增加 PGRN 的表达，这证实了 REST 对 PGRN 表达的抑制作用。进一步的双荧光素酶报告基因检测也证实了 REST 对 PGRN 启动子活性的抑制作用，表明 REST 可能是调节小胶质细胞样 BV2 细胞迁移的 PGRN 的抑制性转录因子。总之，本研究表明转录因子 REST 可能通过 PGRN 作为小胶质细胞迁移的抑制剂。
