Notch signaling pathway and Cdx2 expression in the development of Barrett's esophagus
Notch signaling pathway and Cdx2 expression in the development of Barrett's esophagus
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DOI：
10.1038/labinvest.2012.56
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发表时间：
2012-06-01
影响因子：
5
通讯作者：
Kinoshita, Yoshikazu
中科院分区：
文献类型：
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作者：
Tamagawa, Yuji;Ishimura, Norihisa;Kinoshita, Yoshikazu
反流胆汁酸诱导的食道干细胞CDX2表达可能是Barrett‘s食道发生发展的重要因素，而Notch信号转导通路是决定细胞分化的重要分子信号通路。Atoh1(一种与Notch信号转导相关的因子)在干细胞向杯状细胞分化的过程中起着重要作用。然而，Notch信号通路和CDX2在Barrett‘s食道发育过程中的表达之间的关系尚未被探索。本研究的目的是探讨食道上皮细胞中Notch信号与CDX2的相互关系。采用实时定量聚合酶链式反应(PCR)和免疫组织化学染色方法，检测伴有杯状细胞(IM+)和不伴有杯状细胞(IM-)的食管肠化生(IM)组织中CDX2、MUC2和Notch1、Hes1、Atoh1的表达。在体外实验中，我们采用了人食道上皮细胞系(OE33、OE19和Het-1A)。应用CDX2表达载体诱导细胞表达CDX2后，用实时定量聚合酶链式反应和免疫印迹分析Notch1、Hes1、Atoh1、CDx2和MUC2的表达变化。分析胆汁酸对细胞Notch1、Hes1、Atoh1、CDx2和MUC2表达的影响。在IM(+)标本中，Hes1表达被抑制，Atoh1和MUC2表达增强。在细胞中强制表达CDX2抑制Hes1，增强Atoh1和MUC2的表达，而胆汁酸抑制Hes1，增强Atoh1，CDX2和MUC2的表达。另一方面，这些效应被基于siRNA的CDX2下调所阻断。胆汁酸刺激增强CDX2的表达可能通过与Notch信号通路相互作用而诱导食管柱细胞的肠道分化。实验室调查(2012年)92896-909；doi：10.1038/Labinvest.2012.56；2012年3月26日在线发布
Cdx2 expression in esophageal stem cells induced by reflux bile acids may be an important factor for development of Barrett's esophagus, whereas Notch signaling is a molecular signaling pathway that plays an important role in the determination of cell differentiation. ATOH1 (a factor associated with Notch signaling) plays an important role in differentiation of stem cells into goblet cells. However, the relationship between the Notch signaling pathway and Cdx2 expression in the development of Barrett's esophagus has not been explored. The aim of this study was to investigate the interrelationship between Notch signaling and Cdx2 in esophageal epithelial cells. The expressions of Cdx2, MUC2, and intracellular signaling molecules related to Notch signaling (Notch1, Hes1, and ATOH1) were examined using real-time polymerase chain reaction (PCR) and immunohistochemical staining with biopsy specimens obtained from esophageal intestinal metaplasia (IM) with goblet cells (IM(+)) and columnar epithelium not accompanied by goblet cells (IM(-)). For in vitro experiments, we employed human esophageal epithelial cell lines (OE33, OE19, and Het-1A). After forced Cdx2 expression by applying a Cdx2 expression vector to the cells, changes in the expressions of Notch1, Hes1, ATOH1, Cdx2, and MUC2 were analyzed by real-time PCR and western blot analysis. Changes in expressions of Notch1, Hes1, ATOH1, Cdx2, and MUC2 in cells were analyzed following stimulation with bile acids in the presence or absence of Cdx2 blocking with Cdx2-siRNA. Suppressed Hes1 and enhanced ATOH1 and MUC2 expressions were identified in IM(+) specimens. Forced expression of Cdx2 in cells suppressed Hes1, and enhanced ATOH1 and MUC2 expressions, whereas bile acids suppressed Hes1, and enhanced ATOH1, Cdx2, and MUC2 expressions. On the other hand, these effects were blocked by siRNA-based Cdx2 downregulation. Enhanced expression of Cdx2 by stimulation with bile acids may induce intestinal differentiation of esophageal columnar cells by interaction with the Notch signaling pathway. Laboratory Investigation (2012) 92, 896-909; doi:10.1038/labinvest.2012.56; published online 26 March 2012