Aberrant Reduction of MiR-141 Increased CD47/CUL3 in Hirschsprung's Disease
Aberrant Reduction of MiR-141 Increased CD47/CUL3 in Hirschsprung's Disease
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DOI：
10.1159/000356601
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发表时间：
2013-01-01
影响因子：
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通讯作者：
Xu, Xiaoqun
中科院分区：
文献类型：
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作者：
Tang, Weibing;Qin, Jingjing;Xu, Xiaoqun
背景：MiR-141已被证实与多种人类疾病相关。然而，miR-141是否参与先天性巨结肠症（HSCR）的发病机制仍不清楚。在这里，我们设计实验来揭示 miR-141 和 HSCR 之间的关系。方法：采用实时定量PCR和Western blot检测70例HSCR组织与60例对照组织中miR-141及其潜在基因的表达水平。应用亚硫酸氢盐测序PCR（BSP）分析来解释miR-141异常表达水平的可能机制。我们采用双荧光素酶报告基因测定来验证 miR-141 和 CD47/CUL3 之间的调节关系。分别通过Transwell实验、MTT实验和流式细胞术检测细胞迁移、增殖、凋亡和细胞周期进展。结果：与对照组相比，HSCR患者结肠组织中miR-141表达下调，CD47和CUL3表达升高，miR-141诱导的CD47和CUL3水平升高降低了293T和SH-SY5Y细胞的增殖和迁移。此外，这种抑制可通过减少 CD47 和 CUL3 来逆转。在 HSCR 组织中证实了 miR-141 基因启动子区 CpG 岛的高甲基化。结论：miR-141的异常减少可能在HSCR的发病机制中发挥重要作用，并抑制细胞迁移和增殖能力。本研究首次证实了miR-141及其靶基因在HSCR发生中的作用，为我们研究先天性巨结肠的发病机制提供了新的方向。 (C) 2013 S. Karger AG，巴塞尔
Background: MiR-141 has been confirmed to be associated with various human diseases. However, whether miR-141 is involved in the pathogenesis of Hirschsprung's disease (HSCR) remains unknown. Here, we design the experiment to reveal the relationship between miR-141 and HSCR. Methods: Quantitative real-time PCR and Western blot were used to detect the expression levels of miR-141 and its potential genes in 70 tissues of HSCR compared with 60 controls. Bisulfite sequencing PCR (BSP) assay was applied to explain the possible mechanism of the aberrant expression level of miR-141. We employed a dual-luciferase reporter assay to validate the regulation relation between miR-141 and CD47/CUL3. Cell migration, proliferation, apoptosis, and cell cycle progression were examined by transwell assay, MTT assay, and flow cytometry, respectively. Results: MiR-141 was down-regulated whereas CD47 and CUL3 expression was increased in colon tissues from patients with HSCR compared with control group, The increased level of CD47 and CUL3 induced by miR-141 reduced proliferation and migration of 293T and SH-SY5Y cells. Furthermore, this suppression was reversed by reducing of CD47 and CUL3. Hypermethylation of a CpG Island in the promoter region of miR-141 gene was confirmed in HSCR tissues. Conclusion: Aberrant reduction of miR-141 may play an important role in the pathogenesis of HSCR with the inhibiting affection on cell migration and proliferation abilities. The present study demonstrates for the first time the role of miR-141 and its target genes in the occurrence of HSCR, and provides us a new direction for the study of the pathogenesis of Hirschsprung's disease. (C) 2013 S. Karger AG, Basel