SLC35A2 Deficiency Promotes an Epithelial-to-Mesenchymal Transition-like Phenotype in Madin-Darby Canine Kidney Cells.
SLC35A2 Deficiency Promotes an Epithelial-to-Mesenchymal Transition-like Phenotype in Madin-Darby Canine Kidney Cells.
复制标题
SLC35A2缺乏促进Madin-Darby犬肾细胞上皮向间充质转化样表型。
DOI：
10.3390/cells11152273
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发表时间：
2022-07-23
期刊：
影响因子：
6
通讯作者：
Maszczak-Seneczko, Dorota
中科院分区：
文献类型：
--
作者：
Kot, Magdalena;Mazurkiewicz, Ewa;Wiktor, Maciej;Wiertelak, Wojciech;Mazur, Antonina Joanna;Rahalevich, Andrei;Olczak, Mariusz;Maszczak-Seneczko, Dorota
关键词：
在哺乳动物细胞中，SLC35A2为半乳糖化反应提供UDP-半乳糖，该反应主要发生在高尔基体腔内。相应基因的突变会导致一种先天性糖基化障碍(SLC35A2-CDG)的亚型。虽然越来越多的患者被诊断为SLC35A2-CDG，但半乳糖化缺陷与疾病症状之间的联系尚不完全清楚。据报道，糖基化受损可能触发上皮间充质转化(EMT)的过程。因此，我们研究了SLC35A2活性的丧失是否会促进非恶性上皮细胞系的EMT。为此，我们在Madin-Darby犬肾(MDCK)细胞中敲除了SLC35A2基因。所产生的克隆采用了细长的纺锤形形态，并显示出细胞与细胞之间的粘附力受损。利用qPCR和Western blotting，我们发现E-钙粘蛋白在基因敲除中下调，而纤维连接蛋白和波形蛋白水平升高。此外，基因敲除的细胞表现出波形蛋白中间丝的重组，并改变了波形蛋白结合蛋白甲酰亚氨基转移酶环脱氨酶(FTCD)的亚细胞分布。此外，SLC35A2的枯竭引发了高尔基体压实。最后，SLC35A2基因敲除显示出更强的运动性和侵袭性。总之，SLC35A2缺陷的MDCK细胞表现出EMT的几个特征。我们的发现指出了SLC35A2作为上皮表型的守门人的新角色。
In mammalian cells, SLC35A2 delivers UDP–galactose for galactosylation reactions that take place predominantly in the Golgi lumen. Mutations in the corresponding gene cause a subtype of a congenital disorder of glycosylation (SLC35A2-CDG). Although more and more patients are diagnosed with SLC35A2-CDG, the link between defective galactosylation and disease symptoms is not fully understood. According to a number of reports, impaired glycosylation may trigger the process of epithelial-to-mesenchymal transition (EMT). We therefore examined whether the loss of SLC35A2 activity would promote EMT in a non-malignant epithelial cell line. For this purpose, we knocked out the SLC35A2 gene in Madin–Darby canine kidney (MDCK) cells. The resulting clones adopted an elongated, spindle-shaped morphology and showed impaired cell–cell adhesion. Using qPCR and western blotting, we revealed down-regulation of E-cadherin in the knockouts, while the fibronectin and vimentin levels were elevated. Moreover, the knockout cells displayed reorganization of vimentin intermediate filaments and altered subcellular distribution of a vimentin-binding protein, formiminotransferase cyclodeaminase (FTCD). Furthermore, depletion of SLC35A2 triggered Golgi compaction. Finally, the SLC35A2 knockouts displayed increased motility and invasiveness. In conclusion, SLC35A2-deficient MDCK cells showed several hallmarks of EMT. Our findings point to a novel role for SLC35A2 as a gatekeeper of the epithelial phenotype.
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影响因子：
4.6
作者：
Malek N;Michrowska A;Mazurkiewicz E;Mrówczyńska E;Mackiewicz P;Mazur AJ
通讯作者：
Mazur AJ
影响因子：
3
作者：
Maszczak-Seneczko, Dorota;Olczak, Teresa;Wunderlich, Livius;Olczak, Mariusz
通讯作者：
Olczak, Mariusz
影响因子：
3.5
作者：
Maszczak-Seneczko, Dorota;Sosicka, Paulina;Olczak, Mariusz
通讯作者：
Olczak, Mariusz
影响因子：
3.7
作者：
Bazan B;Wiktor M;Maszczak-Seneczko D;Olczak T;Kaczmarek B;Olczak M
通讯作者：
Olczak M
影响因子：
4.8
作者：
Ju, TZ;Brewer, K;Canfield, WM
通讯作者：
Canfield, WM