Center for Stem Cell and Translational Medicine, School of Life Sciences, Anhui University, Hefei 230601, Anhui, China.
Department of Biostatistics, School of Life Sciences, Anhui University, Hefei 230601, Anhui, China.
Aging (Albany NY). 2021 Mar 24;13(6):9011-9027. doi: 10.18632/aging.202749.
Protein inhibitor of activated STAT1 (PIAS1), a small ubiquitin-like modifier (SUMO) E3 ligase, was considered to be an inhibitor of STAT1 by inhibiting the DNA-binding activity of STAT1 and blocking STAT1-mediated gene transcription in response to cytokine stimulation. PIAS1 has been determined to be involved in modulating several biological processes such as cell proliferation, DNA damage responses, and inflammatory responses, both and . However, the role played by PIAS1 in regulating neurodegenerative diseases, including Alzheimer's disease (AD), has not been determined. In our study, significantly different expression levels of PIAS1 between normal controls and AD patients were detected in four regions of the human brain. Based on a functional analysis of in undifferentiated mouse hippocampal neuronal HT-22 cells, we observed that the expression levels of several AD marker genes could be inhibited by overexpression. Moreover, the proliferation ability of HT-22 cells could be promoted by the overexpression of . Furthermore, we performed RNA sequencing (RNA-seq) to evaluate and quantify the gene expression profiles in response to overexpression in HT-22 cells. As a result, 285 significantly dysregulated genes, including 79 upregulated genes and 206 downregulated genes, were identified by the comparison of /+ cells with WT cells. Among these genes, five overlapping genes, including early growth response 1 (), early growth response 2 (), early growth response 3 (), FBJ osteosarcoma oncogene () and fos-like antigen 1 (), were identified by comparison of the transcription factor binding site (TFBS) prediction results for STAT1, whose expression was evaluated by qPCR. Three cell cycle inhibitors, p53, p18 and p21, were significantly downregulated with the overexpression of . Analysis of functional enrichment and expression levels showed that basic region leucine zipper domain-containing transcription factors including zinc finger C2H2 (zf-C2H2), homeobox and basic/helix-loop-helix (bHLH) in several signaling pathways were significantly involved in PIAS1 regulation in HT-22 cells. A reconstructed regulatory network under PIAS1 overexpression demonstrated that there were 43 related proteins, notably Nr3c2, that directly interacted with PIAS1.
蛋白质抑制剂激活 STAT1（PIAS1）是一种小泛素样修饰物（SUMO）E3 连接酶，被认为是 STAT1 的抑制剂，通过抑制 STAT1 的 DNA 结合活性并阻断细胞因子刺激下的 STAT1 介导的基因转录。PIAS1 已被确定参与调节多种生物学过程，如细胞增殖、DNA 损伤反应和炎症反应，既 又 。然而，PIAS1 在调节神经退行性疾病，包括阿尔茨海默病（AD）中的作用尚未确定。在我们的研究中，在人类大脑的四个区域检测到正常对照和 AD 患者之间 PIAS1 的表达水平存在显著差异。基于对未分化的小鼠海马神经元 HT-22 细胞中的功能分析，我们观察到几种 AD 标志物基因的表达水平可以被 过表达抑制。此外，PIAS1 的过表达可以促进 HT-22 细胞的增殖能力。此外，我们进行了 RNA 测序（RNA-seq），以评估和量化 HT-22 细胞中 过表达的基因表达谱。结果，与 WT 细胞相比，在 /+细胞中鉴定出 285 个显著失调的基因，包括 79 个上调基因和 206 个下调基因。在这些基因中，通过比较 STAT1 的转录因子结合位点（TFBS）预测结果，鉴定出五个重叠基因，包括早期生长反应 1（）、早期生长反应 2（）、早期生长反应 3（）、FBJ 骨肉瘤癌基因（）和 fos 样抗原 1（）。通过 qPCR 评估其表达。三个细胞周期抑制剂，p53、p18 和 p21，随着 PIAS1 的过表达而显著下调。功能富集和表达水平分析表明，锌指 C2H2（zf-C2H2）、同源盒和碱性/螺旋-环-螺旋（bHLH）等几种信号通路中的基本区亮氨酸拉链域转录因子包括在内，在 HT-22 细胞中 PIAS1 的调节中显着参与。在 PIAS1 过表达下重建的调控网络表明，有 43 种相关蛋白，特别是 Nr3c2，与 PIAS1 直接相互作用。
