A novel regulatory network among LncRpa, CircRar1, MiR‑671 and apoptotic genes promotes lead‑induced neuronal cell apoptosis
发布时间 :2016-09-20
研究对象:小鼠神经细胞、lncRNA、circRNA、miRNA 期刊:Archives of Toxicology 影响因子:6.637 合作单位:广州医科大学 发表时间:2016年9月

摘要

       Lead is a metal that has toxic effects on the developing nervous system. However, the mechanisms underlying lead-induced neurotoxicity are not well understood. Non-coding RNAs (ncRNAs) play an important role in epigenetic regulation, but few studies have examined the function of ncRNAs in lead-induced neurotoxicity. We addressed this in the present study by evaluating the functions of a long non-coding RNA (named lncRpa) and a circular RNA (named circRar1) in a mouse model of leadinduced neurotoxicity. High-throughput RNA sequencing showed that both lncRpa and circRar1 promoted neuronal apoptosis. We also found that lncRpa and circRar1 induced the upregulation of apoptosis-associated factors caspase8 and p38 at the mRNA and protein levels via modulation of their common target microRNA miR-671. This is the first report of a regulatory interaction among a lncRNA, circRNA, and miRNA mediating neuronal apoptosis in response to lead toxicity.

关键词:CircRNA、LncRNA、MiRNA、Cell apoptosis、Lead Neurotoxicity

研究背景

  铅作为一种神经发育性强烈毒药,会导致神经性坏死。然而,关于铅致神经中毒机理目前并没有很好的解释。非编码RNAncRNAs表观遗传调控领扮演者重要的角色,但是,ncRNA参与铅致神经中毒的功能研究却很少,本研究突出了lncRNAs、circRNAs和miRNAs的相互作用旨在解释ncRNAs在中毒神经细胞凋亡过程中的调控机制

方法流程


部分研究结果

1. LncRpa circRar1促进细胞凋亡

  分别过表达和干扰lncRpa、circRar1,检测过表达和RNA干扰效率,利用FCMTUNEL进行凋亡检测,同时对凋亡相关蛋白进行表达检测。

2. LncRpa、circRar1直接互作miR‑671

  首先利用数据库预测lncRpacircRar1共同互作的miRNA,荧光原位杂交(FISH进行lncRpa、circRar1定位,再开展荧光素酶报告基因和RAP-qPCR进行验证。结显示lncRpacircRar1能够与miR‑671直接互作。

3. LncRpacircRar1调控miR‑671的表达

  荧光原位杂交结果显示lncRpacircRar1、miR‑671N2a细胞中共表达。相互之间开展过表达和RNA干扰实验结果显示lncRpacircRar1相互促进;miR‑671与lncRpa/circRar1之间则是一种负调控的关系


伯信合作技术

CircRNA过表达载体构建、荧光原位杂交(FISHRNA antisense purification(RAP)

参考文献

Aruo Nan, Lijian Chen, Nan Zhang, et al. A novel regulatory network among LncRpa, CircRar1, MiR-671 and apoptotic genes promotes lead-induced neuronal cell apoptosis[J]. 2016.

原文链接

http://link.springer.com/article/10.1007%2Fs00204-016-1837-1

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