Specific N-glycan alterations are coupled in EMT induced by different density cultivation of MCF 10A epithelial cells
Specific N-glycan alterations are coupled in EMT induced by different density cultivation of MCF 10A epithelial cells
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MCF 10A 上皮细胞不同密度培养诱导的 EMT 中耦合了特定的 N-聚糖改变
DOI：
10.1007/s10719-016-9754-3
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发表时间：
2017
影响因子：
3
通讯作者：
Song Linsheng
中科院分区：
文献类型：
--
作者：
Xu Qingsong;Niu Xueming;Wang Wenjing;Yang Wen;Du Yuguang;Gu Jianguo;Song Linsheng
上皮-间质转化（EMT）是肿瘤进展中的一个过程，在此过程中，癌细胞经历戏剧性的变化，获得高度侵袭性的特性。在广泛采用的TGF-β诱导的EMT模型中，我们先前观察到N-聚糖上的二等分GlcNAc的表达显著降低。在此，我们用MCF 10A细胞系进行了体外研究。MCF 10A细胞在低密度下自发发生形态学和表型EMT样改变，包括细胞片边缘延长、纺锤体形状延长、细胞骨架重组、波形蛋白和纤连蛋白表达上调、连环蛋白重新分布和钙粘蛋白转换。此外，这些表型变化与特异性N-聚糖改变相关。有趣的是，在稀疏细胞条件下诱导的EMT过程中，二等分GlcNAc结构的数量减少，相反，β1-6 GlcNAc分支的形成明显上调。我们进一步研究了β1-整合素上的N-聚糖，β1-整合素是一些糖基转移酶的良好靶点。与E4-PHA凝集素的反应性降低，而L4-PHA凝集素，它承认分支GlcNAc，染色增加在稀疏的细胞条件下相比，密集的细胞条件。综上所述，这些结果表明，特异性N-聚糖改变耦合在EMT过程中，并在低细胞密度下促进细胞迁移。
Epithelial-mesenchymal transition (EMT) is a process in tumor progression during which cancer cells undergo dramatic changes acquiring highly invasive properties. In a widespread adoption of TGF-β-induced EMT model, we have previously observed that expression of bisecting GlcNAc onN-glycans was dramatically decreased. Herein, we performedin vitrostudies with the MCF10A cell line. In response to low cell density, MCF10A cells suffered spontaneously morphologic and phenotypic EMT-like changes, including elongated spindle shape, extended out from edge of the cell sheet, cytoskeleton reorganization, vimentin and fibronectin up-regulation, catenins redistribution, and cadherin switching. Moreover, these phenotypic changes were associated with specificN-glycan alterations. Interestingly, the amounts of bisecting GlcNAc structure were declined, by contrast, the formation of β1-6 GlcNAc branches were obviously up-regulated during the EMT induced by sparse cell conditions. We further investigatedN-glycans on the β1-integrin, which is a good target of some glycosyltransferases. The reactivity with E4-PHA lectin decreased, whereas the staining for L4-PHA lectin, which recognizes branched GlcNAc, increased in sparse cell conditions compared with dense cell conditions. Taken together, these results demonstrated that specificN-glycan alterations are coupled in EMT process and promoted cells migration at a low cell density.