Podoplanin-mediated TGF-β-induced epithelial-mesenchymal transition and its correlation with bHLH transcription factor DEC in TE-11 cells.
Podoplanin-mediated TGF-β-induced epithelial-mesenchymal transition and its correlation with bHLH transcription factor DEC in TE-11 cells.
复制标题
Podopplanin介导的TGF-β诱导的上皮 - 间质转变及其与TE-11细胞中BHLH转录因子DEC的相关性。
DOI：
10.3892/ijo.2016.3445
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发表时间：
2016-06
影响因子：
5.2
通讯作者：
Kijima H
中科院分区：
文献类型：
--
作者：
Wu Y;Liu Q;Yan X;Kato Y;Tanaka M;Inokuchi S;Yoshizawa T;Morohashi S;Kijima H
据报道，泊多普宁参与了集体细胞侵袭，这是另一种有别于单一细胞侵袭的肿瘤侵袭方式，即所谓的上皮-间充质转化(EMT)。本研究探讨了食管鳞癌组织中泊多普宁与EMT相关标记物的相关性，并探讨了其与碱性螺旋-环-螺旋(BHLH)转录因子分化的胚胎软骨细胞(DEC)1和DEC2的关系。对三种ESCC细胞系和人鳞状细胞癌A431细胞进行podoplan in和EMT标记物的蛋白质印迹分析，以及DEC1和DEC2的表达。RT-qPCR法和Western blotting结果显示，转化生长因子-β可上调足阳素和间充质标志物(如N-钙粘附素和波形蛋白)的表达，降低上皮标志物(如克劳丁-4和E-钙粘附素)的表达，并伴随Smad2的磷酸化和黏附蛋白的激活。此外，转化生长因子-β对DEC1和DEC2的表达有不同的影响，即上调DEC1，下调DEC2。分别用CCK-8比色法、Matrigel侵袭实验和伤口愈合实验进一步分析细胞的增殖、侵袭和迁移能力。与错配siRNA组相比，敲除podoplan in基因的细胞的增殖、侵袭和迁移能力明显丧失。除上述变化外，抗Podoplan in的siRNA还可诱导Claudin-4的表达，但不能诱导Claudin-1或E-cadherin的表达。相反，过表达DEC1和DEC2对泊多拉宁的作用相反，但对Claudin-4的影响很小。这些数据表明，泊多拉宁与TE-11细胞的EMT密切相关，并可能直接或间接地受到bHLH转录因子DEC1和DEC2的调控。
Podoplanin is reported involved in the collective cell invasion, another tumor invasion style which is distinct from the single cell invasion, so-called epithelial-mesenchymal transition (EMT). In this study, we investigated the correlation between podoplanin and EMT-related markers in esophageal squamous cell carcinoma (ESCC), and evaluated its linkage with the basic helix-loop-helix (bHLH) transcription factor differentiated embryonic chondrocyte (DEC) 1 and DEC2. Three ESCC cell lines and human squamous cell carcinoma A431 cells were subjected to western blot analyses for podoplanin and EMT markers, as well as the expression of DEC1 and DEC2. By RT-qPCR and western blotting, we found that TGF-β increased the expression of podoplanin and mensenchymal markers (e.g., N-cadherin and vimentin), while decreased the expression of epithelial markers (e.g., Claudin-4 and E-cadherin), accompanied by Smad2 phosphorylation and slug activation. Moreover, TGF-β has different effects on the expression of DEC1 and DEC2, that is, it upregulates DEC1, but downregulates DEC2. Capability of cell proliferation, invasion and migration were further analyzed using CCK-8 assay, Matrigel-invasion assay, and the wound-healing assay, respectively. The proliferation, invasion and migration ability were significantly lost in podoplanin-knockdown cells when compared with the scrambled siRNA group. In addition to these changes, the expression of Claudin-4, but not that of Claudin-1 or E-cadherin, was induced by the siRNA against podoplanin. On the contrary, overexpression of DEC1 and DEC2 exhibits opposite effects on podoplanin, but only slight effect on Claudin-4 was detected. These data indicated that podoplanin is significantly associated with EMT of TE-11 cells, and may be directly or indirectly regulated by bHLH transcription factors DEC1 and DEC2.