Altered expression of microRNAs in the response to ER stress
Altered expression of microRNAs in the response to ER stress
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内质网应激反应中 microRNA 表达的改变
DOI：
10.1007/s11434-014-0657-z
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发表时间：
2015-01-01
期刊：
影响因子：
18.9
通讯作者：
Li, Zhaoyong
中科院分区：
文献类型：
--
作者：
Dai, Limin;Huang, Chuan;Li, Zhaoyong
MicroRNAs是一类非编码的小RNA，在多种生物和病理过程中发挥着关键作用。内质网应激是由内质网管腔内未折叠或错误折叠的蛋白质积聚引起的，由各种生理事件和病理损伤触发。在这里，我们使用RNA深度测序分析发现，在内质网应激下，HeLa和HEK293细胞中一些microRNAs的表达发生了变化。内质网应激细胞中DgCr8、DROSHA、Exportin-5、Dice和Ago2的蛋白质和RNA水平没有明显变化，这表明microRNA表达的变化可能不是由microRNA生物发生途径引起的，而是由其他未知因素引起的。实时荧光定量PCR检测证实，ER胁迫下HeLa和HEK293细胞中hsa-miR-423-5p表达上调，hsa-miR-221-3p和hsa-miR-452-5p表达下调。荧光素酶活性和Western印迹分析证实CDKN1A是hsa-miR-423-5p的直接靶标，CDKN1B是hsa-miR-221-3p和hsa-miR-452-5p的直接靶标。我们推测，通过调节它们的靶标，microRNAs可能在对内质网应激的适应性反应中协同发挥调节作用。
MicroRNAs, a class of small noncoding RNAs, play key roles in diverse biological and pathological processes. ER stress, resulting from the accumulation of unfolded or misfolded proteins in the ER lumen, is triggered by various physiological events and pathological insults. Here, using RNA deep sequencing analysis, we found that the expression of some microRNAs was altered in HeLa and HEK293 cells under ER stress. Protein and RNA levels of DGCR8, Drosha, Exportin-5, Dicer, and Ago2 showed no significant alteration in ER-stressed cells, which suggested that the change in microRNA expression might not be caused by the microRNA biogenesis pathway but by other, unknown factors. Real-time PCR assays confirmed that hsa-miR-423-5p was up-regulated, whereas hsa-miR-221-3p and hsa-miR-452-5p were down-regulated, in both HeLa and HEK293 cells under ER stress. Luciferase activity and Western blot assays verified that CDKN1A was a direct target of hsa-miR-423-5p and that CDKN1B was a direct target of hsa-miR-221-3p and hsa-miR-452-5p. We speculated that by regulating their targets, microRNAs might function cooperatively as regulators in the adaptive response to ER stress.