长链非编码 RNA EPB41L4A-AS1 通过介导 HDAC2 的核仁易位来调节糖酵解和谷氨酰胺分解。
LncRNA EPB41L4A-AS1 regulates glycolysis and glutaminolysis by mediating nucleolar translocation of HDAC2.
机构信息
School of Life Sciences, Tsinghua University, Beijing 100084, PR China; Key Lab in Healthy Science and Technology, Division of Life Science, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, PR China.
State Key Laboratory of Chemical Oncogenomics, Graduate School at Shenzhen, Tsinghua University, Shenzhen, PR China; Key Lab in Healthy Science and Technology, Division of Life Science, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, PR China; Open FIESTA Center, Tsinghua University, Shenzhen 518055, PR China.
出版信息
EBioMedicine. 2019 Mar;41:200-213. doi: 10.1016/j.ebiom.2019.01.035. Epub 2019 Feb 19.
BACKGROUND
LncRNAs have been found to be involved in various aspects of biological processes. In this study, we aimed to uncover the molecular mechanisms of lncRNA EPB41L4A-AS1 in regulating glycolysis and glutaminolysis in cancer cells.
METHODS
The expression of EPB41L4A-AS1 in cancer patients was analyzed in TCGA and GEO datasets. The level of cellular metabolism was determined by extracellular flux analyzer. The relationship between p53 and EPB41L4A-AS1 was explored by qRT-PCR, luciferase assay and ChIP assay. The interactions between EPB41L4A-AS1 and HDAC2 or NPM1 were determined by RNA immunoprecipitation, RNA pull-down assay and RNA-FISH- immunofluorescence.
FINDINGS
EPB41L4A-AS1 was a p53-regulated gene. Low expression and deletion of lncRNA EPB41L4A-AS1 were found in a variety of human cancers and associated with poor prognosis of cancer patients. Knock down EPB41L4A-AS1 expression triggered Warburg effect, demonstrated as increased aerobic glycolysis and glutaminolysis. EPB41L4A-AS1 interacted and colocalized with HDAC2 and NPM1 in nucleolus. Silencing EPB41L4A-AS1 reduced the interaction between HDAC2 and NPM1, released HDAC2 from nucleolus and increased its distribution in nucleoplasm, enhanced HDAC2 occupation on VHL and VDAC1 promoter regions, and finally accelerated glycolysis and glutaminolysis. Depletion of EPB41L4A-AS1 increased the sensitivity of tumor to glutaminase inhibitor in tumor therapy.
INTERPRETATION
EPB41L4A-AS1 functions as a repressor of the Warburg effect and plays important roles in metabolic reprogramming of cancer.
背景
长链非编码 RNA（lncRNA）已被发现参与生物过程的各个方面。在这项研究中，我们旨在揭示 lncRNA EPB41L4A-AS1 调节癌细胞糖酵解和谷氨酰胺分解的分子机制。
方法
在 TCGA 和 GEO 数据集分析癌症患者中 EPB41L4A-AS1 的表达。通过细胞外通量分析器测定细胞代谢水平。通过 qRT-PCR、荧光素酶报告基因检测和 ChIP 实验探究 p53 与 EPB41L4A-AS1 的关系。通过 RNA 免疫沉淀、RNA 下拉实验和 RNA-FISH-免疫荧光实验确定 EPB41L4A-AS1 与 HDAC2 或 NPM1 的相互作用。
发现
EPB41L4A-AS1 是 p53 调控的基因。在多种人类癌症中发现 lncRNA EPB41L4A-AS1 表达水平降低或缺失，且与癌症患者的不良预后相关。敲低 EPB41L4A-AS1 表达会引发瓦博格效应，表现为有氧糖酵解和谷氨酰胺分解增加。EPB41L4A-AS1 与核仁中的 HDAC2 和 NPM1 相互作用并共定位。沉默 EPB41L4A-AS1 减少了 HDAC2 和 NPM1 之间的相互作用，使 HDAC2 从核仁中释放出来并增加其在核质中的分布，增强了 HDAC2 在 VHL 和 VDAC1 启动子区域的占据，最终加速糖酵解和谷氨酰胺分解。EPB41L4A-AS1 的耗竭增加了肿瘤治疗中谷氨酰胺酶抑制剂对肿瘤的敏感性。
解释
EPB41L4A-AS1 作为瓦博格效应的抑制剂发挥作用，在肿瘤代谢重编程中发挥重要作用。
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