Laboratory of Glycobiology, The Noguchi Institute, 1-9-7 Kaga, Itabashi, Tokyo, 173-0003, Japan.
Laboratory of Glycobiology, Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata, 940-2188, Japan.
Histochem Cell Biol. 2020 Jan;153(1):17-26. doi: 10.1007/s00418-019-01822-3. Epub 2019 Oct 12.
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.
我们之前已经证明,在 MDA-MB-231 人乳腺癌细胞表面糖蛋白的 N-糖链上表达二糖 GalNAcβ1-4GlcNAc(LacdiNAc),可抑制其恶性特性,如裸鼠肿瘤形成。在这里,我们报告了过表达β4-N-乙酰半乳糖胺基转移酶 4 的 MDA-MB-231 细胞两种克隆细胞的形态外观和黏附特性的变化。与模拟转染细胞和原始 MDA-MB-231 细胞的纺锤形相比,克隆细胞呈现出鹅卵石状。这与细胞表面 E-钙黏蛋白(上皮细胞标志物)的表达增加和 N-钙黏蛋白、波形蛋白、α-平滑肌肌动蛋白和 ZEB1(间充质细胞标志物)的表达减少有关。此外,与模拟转染细胞相比,克隆细胞的划痕愈合试验迁移活性较低。这些结果表明,这些克隆细胞可能正在发生间质-上皮转化。此外,还观察到对细胞外基质蛋白(如纤连蛋白、I 型胶原、IV 型胶原和层粘连蛋白)的黏附增加。克隆细胞铺展和增大,而模拟转染细胞在缺乏胎牛血清的情况下在层粘连蛋白包被的平板上表现出较差的铺展,表明细胞表面糖蛋白上 LacdiNAc 的表达导致细胞黏附和铺展特性的变化,特别是对层粘连蛋白。