We demonstrated previously that the expression of the disaccharide, GalNAcβ1 → 4GlcNAc (LacdiNAc), on N-glycans of cell surface glycoproteins in MDA-MB-231 human breast cancer cells suppresses their malignant properties such as tumor formation in nude mice. Here, we report changes in the morphological appearance and adhesive properties of two kinds of clonal cells of MDA-MB-231 cells overexpressing β4-N-acetyl-galactosaminyltransferase 4. The clonal cells exhibited a cobble stone-like shape as compared to a spindle-like shape of the mock-transfected cells and the original MDA-MB-231 cells. This was associated with an increased expression of cell surface E-cadherin, a marker of epithelial cells, and a decreased expression of N-cadherin, vimentin, α-smooth muscle actin and ZEB1, markers of mesenchymal cells. In addition, the clonal cells showed a lower migratory activity compared to the mock-transfected cells by wound-healing assay. These results suggest that mesenchymal-epithelial transition may be occurring in these clonal cells. Furthermore, increased adhesion to extracellular matrix proteins such as fibronectin, collagen type I, collagen type IV, and laminin was observed. The clonal cells spread and enlarged, whereas the mock-transfected cells demonstrated poor spreading on laminin-coated plates in the absence of fetal calf serum, indicating that expression of LacdiNAc on cell surface glycoproteins results in changes in cell adhesive and spreading properties particularly to laminin.

中文翻译：

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MDA-MB-231人乳腺癌细胞中N-聚糖的LacdiNAcylation导致细胞形态和粘附特性的变化。

先前我们证明了MDA-MB-231人乳腺癌细胞中细胞表面糖蛋白N聚糖上二糖GalNAcβ1→4GlcNAc（LacdiNAc）的表达抑制了其恶性特性，例如裸鼠的肿瘤形成。在这里，我们报道了两种过度表达β4-N-乙酰基-半乳糖胺基转移酶4的MDA-MB-231细胞的克隆细胞形态和粘附特性的变化。与纺锤体相比，该克隆细胞表现出卵石状的形状。转染的细胞和原始MDA-MB-231细胞具有类似的形状。这与上皮细胞标志物细胞表面E-钙粘蛋白的表达增加以及间充质细胞标志物N-钙粘蛋白，波形蛋白，α平滑肌肌动蛋白和ZEB1的表达减少有关。此外，通过伤口愈合分析，与模拟转染的细胞相比，克隆细胞显示出较低的迁移活性。这些结果表明，在这些克隆细胞中可能发生间充质-上皮转化。此外，观察到与细胞外基质蛋白（如纤连蛋白，I型胶原蛋白，IV型胶原蛋白和层粘连蛋白）的粘附性增加。克隆细胞扩散并扩大，而在没有胎牛血清的情况下，模拟转染的细胞在层粘连蛋白包被的平板上显示出较差的扩散，这表明LacdiNAc在细胞表面糖蛋白上的表达导致细胞黏附和分布特性的改变，特别是层粘连蛋白。这些结果表明，在这些克隆细胞中可能发生间充质-上皮转化。此外，观察到与细胞外基质蛋白（如纤连蛋白，I型胶原蛋白，IV型胶原蛋白和层粘连蛋白）的粘附性增加。克隆细胞扩散并扩大，而在没有胎牛血清的情况下，模拟转染的细胞在层粘连蛋白包被的平板上显示出较差的扩散，这表明LacdiNAc在细胞表面糖蛋白上的表达导致细胞黏附和分布特性的改变，特别是层粘连蛋白。这些结果表明，在这些克隆细胞中可能发生间充质-上皮转化。此外，观察到与细胞外基质蛋白（如纤连蛋白，I型胶原蛋白，IV型胶原蛋白和层粘连蛋白）的粘附性增加。克隆细胞扩散并扩大，而在没有胎牛血清的情况下，模拟转染的细胞在层粘连蛋白包被的平板上显示出较差的扩散，这表明LacdiNAc在细胞表面糖蛋白上的表达导致细胞黏附和分布特性的改变，特别是层粘连蛋白。