NOTCH1 and NOTCH3 coordinate esophageal squamous differentiation through a CSL-dependent transcriptional network.
NOTCH1 and NOTCH3 coordinate esophageal squamous differentiation through a CSL-dependent transcriptional network.
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DOI：
10.1053/j.gastro.2010.08.040
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发表时间：
2010-12
期刊：
影响因子：
29.4
通讯作者：
Nakagawa H
中科院分区：
文献类型：
--
作者：
Ohashi S;Natsuizaka M;Yashiro-Ohtani Y;Kalman RA;Nakagawa M;Wu L;Klein-Szanto AJ;Herlyn M;Diehl JA;Katz JP;Pear WS;Seykora JT;Nakagawa H
Notch受体家族通过细胞间的通讯调节细胞命运。Csl(Cbf-1/RBP-jκ，SU(H)，Lag-1)在造血系统、肠、胰腺和皮肤的细胞谱系指定、分化和增殖过程中驱动规范的Notch介导的基因转录。然而，Notch在食道鳞状上皮生物学中的功能作用仍不清楚。用氯化钙刺激正常食道角质形成细胞诱导其终末分化。鳞状上皮在器官型三维培养中重建，这是人类组织工程的一种形式。在γ分泌酶抑制剂或显性负性主脑样蛋白1(DNMAML1)的培养中，Notch被抑制。通过体外功能获得实验和功能丧失实验评价Notch受体的作用。此外，DNMAML1通过细胞角蛋白K14启动子驱动的Cre(K14Cre)重组Lox-Stop-Lox-DNMAML1靶向小鼠食道。采用报告基因转染法、染色质免疫沉淀(ChIP)法、定量逆转录聚合酶链式反应(RT-PCR)、Western blotting、免疫荧光和免疫组织化学等方法检测Noch调控基因的表达。NOTCH1(N1)在食道鳞状细胞分化开始时被激活。N1(ICN1)的胞内区直接激活NOTCH3(N3)转录，以CSL依赖的方式诱导HES5和早期分化标记物，如总蛋白(IVL)和细胞角蛋白CK13。N3增强了ICN1的活性，是鳞状细胞分化所必需的。K14Cre；DNMAML1小鼠Notch信号的丢失扰乱了食道鳞状细胞的分化，导致N3丢失和基底细胞增生。Notch信号对食道上皮细胞的动态平衡非常重要。特别是，N3和N1在分化过程中的串扰为Notch信号和鳞状上皮生物学提供了新的、机械性的见解。
The Notch receptor family regulates cell fate through cell-cell communication. CSL (CBF-1/RBP-jκ, Su(H), Lag-1) drives canonical Notch-mediated gene transcription during cell lineage specification, differentiation and proliferation in the hematopoietic system, the intestine, the pancreas and the skin. However, the functional roles of Notch in esophageal squamous epithelial biology remain unknown. Normal esophageal keratinocytes were stimulated with calcium chloride to induce terminal differentiation. The squamous epithelia were reconstituted in organotypic three-dimensional culture, a form of human tissue engineering. Notch was inhibited in culture with a γ-secretase inhibitor or dominant negative mastermind-like1 (DNMAML1). The roles of Notch receptors were evaluated by in vitro gain-of-function and loss-of-function experiments. Additionally, DNMAML1 was targeted to the mouse esophagus by cytokeratin K14 promoter-driven Cre (K14Cre) recombination of Lox-STOP-Lox-DNMAML1. Notch-regulated gene expression was determined by reporter transfection, chromatin immunoprecipitation (ChIP) assays, quantitative reverse-transcription polymerase chain reactions (RT-PCR), Western blotting, immunofluorescence and immunohistochemistry. NOTCH1 (N1) was activated at the onset of squamous differentiation in the esophagus. Intracellular domain of N1 (ICN1) directly activated NOTCH3 (N3) transcription, inducing HES5 and early differentiation markers such as involucrin (IVL) and cytokeratin CK13 in a CSL-dependent fashion. N3 enhanced ICN1 activity and was required for squamous differentiation. Loss of Notch signaling in K14Cre;DNMAML1 mice perturbed esophageal squamous differentiation and resulted in N3 loss and basal cell hyperplasia. Notch signaling is important for esophageal epithelial homeostasis. In particular, the crosstalk of N3 with N1 during differentiation provides novel, mechanistic insights into Notch signaling and squamous epithelial biology.
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影响因子：
21.3
作者：
Kolev, Vihren;Mandinova, Anna;Guinea-Viniegra, Juan;Hu, Bing;Lefort, Karine;Lambertini, Chiara;Neel, Victor;Dummer, Reinhard;Wagner, Erwin F.;Dotto, G. Paolo
通讯作者：
Dotto, G. Paolo
影响因子：
11.2
作者：
Ohashi S;Natsuizaka M;Wong GS;Michaylira CZ;Grugan KD;Stairs DB;Kalabis J;Vega ME;Kalman RA;Nakagawa M;Klein-Szanto AJ;Herlyn M;Diehl JA;Rustgi AK;Nakagawa H
通讯作者：
Nakagawa H
影响因子：
4.7
作者：
Lee, James J.;Natsuizaka, Mitsuteru;Nakagawa, Hiroshi
通讯作者：
Nakagawa, Hiroshi
影响因子：
4.3
作者：
Menke, Vivianda;van Es, Johan H.;Clevers, Hans
通讯作者：
Clevers, Hans
影响因子：
10.5
作者：
Nguyen, BC;Lefort, K;Dotto, GP
通讯作者：
Dotto, GP