Transcriptional Profiling Identifies TNS4 Function in Epithelial Tubulogenesis
Transcriptional Profiling Identifies TNS4 Function in Epithelial Tubulogenesis
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DOI：
10.1016/j.cub.2010.12.037
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发表时间：
2011-01-25
期刊：
影响因子：
9.2
通讯作者：
Mostov, Keith E.
中科院分区：
文献类型：
--
作者：
Kwon, Sang-Ho;Nedvetsky, Pavel I.;Mostov, Keith E.
肝细胞生长因子（HGF）在肾小管生成和转移中起着重要作用[1 - 4]。HGF处理在三维培养物中作为囊肿生长的Madin-Darby犬肾（MDCK）细胞诱导肾小管形成[5，6]，其与大多数肾小管形成过程一样，通过不同的中间阶段进行。鉴定与这些阶段相关的基因是理解小管形成的分子机制的核心;然而，由于效率低，不同步的小管形成，分离这些基因是不可行的。在这里，我们开发了一个同步的，高效的tubulogenesis系统，并使用时程转录谱来识别在发育中间体的时间调节基因。tensin 4（TNS4）（一种特别高度上调的基因）的敲低（KD）导致延伸和小管形成的减少，这是小管发生中的两个连续中间体。TNS4的外源性表达标志着上皮层中的侵袭细胞。TNS4的SH2结构域中的突变阻止了在小管形成期间从延伸形成到侵入性迁移的转变，并导致STAT3的基础活化增加。组成型活性STAT3的外源性表达通过突变模拟缺陷。我们的研究强调了TNS4-STAT3轴在上皮层浸润和小管形成中的作用。
Hepatocyte growth factor (HGF) plays central roles in tubulogenesis and metastasis [1-4]. HGF treatment of Madin-Darby canine kidney (MDCK) cells grown as cysts in three-dimensional culture induces tubulogenesis [5, 6], which like most tubulogenic processes proceeds through distinct intermediate phases. Identification of genes associated with these phases is central to understanding the molecular mechanisms of tubulogensis; however, because of inefficient, asynchronous tubule formation, isolating such genes has been unfeasible. Here we developed a synchronous, efficient tubulogenesis system and used time-course transcriptional profiling to identify genes temporally regulated in developmental intermediates. Knockdown (KD) of tensin 4 (TNS4), a particularly highly upregulated gene, leads to a decrease in formation of extensions and tubules, two sequential intermediates in tubulogenesis. Exogenous expression of TNS4 marks invasive cells in an epithelial sheet. A mutation in the SH2 domain of TNS4 prevents the transition from extension formation to invasive migration during tubule formation and leads to increased basal activation of STAT3. Exogenous expression of a constitutively active STAT3 mimics the defect by the mutation. Our study highlights the role of the TNS4-STAT3 axis in epithelial sheet invasion and tubulogenesis.