Knock-down of circular RNA H19 induces human adipose-derived stem cells adipogenic differentiation via a mechanism involving the polypyrimidine tract-binding protein 1
Knock-down of circular RNA H19 induces human adipose-derived stem cells adipogenic differentiation via a mechanism involving the polypyrimidine tract-binding protein 1
复制标题
环状 RNA H19 的敲低通过涉及聚嘧啶束结合蛋白 1 的机制诱导人脂肪干细胞成脂分化
DOI：
10.1016/j.yexcr.2019.111753
复制
发表时间：
2020
影响因子：
3.7
通讯作者：
Hong Li
中科院分区：
文献类型：
--
作者：
Yiyi Zhu;Weiwei Gui;Xihua Lin;Hong Li
目的代谢综合征（MetS）是一组以代谢紊乱为特征的疾病。脂肪组织功能的改变对全身代谢有显著影响，是代谢综合征的关键驱动因素。本研究旨在探讨人环状RNA H19（hsa_circH19）在人脂肪干细胞（hADSCs）中的功能。在hADSC中敲低hsa_circH19后，我们测量了成脂基因的表达。采用油红O、尼罗红染色和甘油三酯测定法检测hsa_circH19在hADSCs分化中的作用。然后，进行RNA Pull-down和RIP测定以探索hsa_circH19的相关RNA结合蛋白。IF进行，以确定潜在的分子调控mechanism.ResultsAfter占混杂因素，高水平的hsa_circH19仍然是一个独立的风险因素MetS。此外，hsa_circH19的敲低显著增加了成脂基因的表达和脂滴的形成。生物信息学分析显示has_circH19与多聚嘧啶片段结合蛋白1（PTBP 1）共享多个结合位点，并且它们的相互作用通过circRNA下拉和RIP测定来验证。结论hsa_circH19基因敲除可通过PTBP 1的靶向作用促进hADCS向脂肪细胞分化。因此，hsa_circH19的表达可能与MetS脂肪组织中的脂质代谢相关。
PurposeThe metabolic syndrome (MetS) is characterized of a cluster of medical disorders. Altered function of adipose tissue has a significant impact on whole-body metabolism and represents a key driver for MetS. In this study, we aim to explore the function of human circular RNA H19 (hsa_circH19) in human adipose-derived stem cells (hADSCs).MethodsThe blood samples from MetS patients and normal subjects were used to determine the expression level of the hsa_circH19. After knock-down of hsa_circH19 in hADSCs, we measured the expression of adipogenic genes. Oil red O, Nile red staining assay and triglyceride assessment were performed to examine the role of hsa_circH19 in hADSCs differentiation. Then, RNA Pull-down and RIP assays were conducted to explore the related RNA binding protein of hsa_circH19. IF was performed to determine the potential molecular regulatory mechanism.ResultsAfter accounting for confounding factors, high levels of hsa_circH19 remained an independent risk factor for MetS. Furthermore, the knockdown of hsa_circH19 significantly increased the expression of adipogenic genes and the formation of lipid droplets. Bioinformatics analyses revealed that has_circH19 shared multiple binding sites with polypyrimidine tract-binding protein 1 (PTBP1) and their interaction was validated by circRNA pull-down and RIP assays. Mechanistically, depletion of hsa_circH19 triggered translocation of sterol-regulatory element binding proteins (SREBP1) from cytoplasm to nucleus in the presence of PTBP1.ConclusionOur experiments suggest that knockdown of hsa_circH19 promotes hADCSs adipogenic differentiation via targeting of PTBP1. In consequence, the expression of hsa_circH19 might correlated to lipid metabolism in adipose tissue from MetS.