Nikolovska Katerina, Spillmann Dorothe, Haier Jörg, Ladányi Andrea, Stock Christian, Seidler Daniela G
Institute of Physiological Chemistry and Pathobiochemistry, University of Münster, Münster, Germany.
Centre for Internal Medicine, Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
PLoS One. 2017 Jan 20;12(1):e0170054. doi: 10.1371/journal.pone.0170054. eCollection 2017.
Although the vast majority of melanomas are characterized by a high metastatic potential, if detected early, melanoma can have a good prognostic outcome. However, once metastasised, the prognosis is bleak. We showed previously that uronyl-2-O sulfotransferase (Ust) and 2-O sulfation of chondroitin/dermatan sulfate (CS/DS) are involved in cell migration. To demonstrate an impact of 2-O sulfation in metastasis we knocked-down Ust in mouse melanoma cells. This significantly reduced the amount of Ust protein and enzyme activity. Furthermore, in vitro cell motility and adhesion were significantly reduced correlating with the decrease of cellular Ust protein. Single cell migration of B16VshUst(16) cells showed a decreased cell movement phenotype. The adhesion of B16V cells to fibronectin depended on α5β1 but not αvβ3 integrin. Inhibition of glycosaminoglycan sulfation or blocking fibroblast growth factor receptor (FgfR) reduced α5 integrin in B16V cell lines. Interestingly, FgfR1 expression and activation was reduced in Ust knock-down cells. In vivo, pulmonary metastasis of B16VshUst cells was prevented due to a reduction of α5 integrin. As a proof of concept UST knock-down in human melanoma cells also showed a reduction in ITGa5 and adhesion. This is the first study showing that Ust, and consequently 2-O sulfation of the low affinity receptor for FgfR CS/DS, reduces Itga5 and leads to an impaired adhesion and migration of melanoma cells.
尽管绝大多数黑色素瘤具有高转移潜能，但如果早期发现，黑色素瘤的预后可能良好。然而，一旦发生转移，预后则不容乐观。我们之前表明，糖醛酸-2-O硫酸转移酶（Ust）以及硫酸软骨素/硫酸皮肤素（CS/DS）的2-O硫酸化参与细胞迁移。为了证明2-O硫酸化在转移中的作用，我们在小鼠黑色素瘤细胞中敲低Ust。这显著降低了Ust蛋白的量和酶活性。此外，体外细胞运动性和黏附性显著降低，这与细胞Ust蛋白的减少相关。B16VshUst(16)细胞的单细胞迁移显示出细胞运动表型降低。B16V细胞与纤连蛋白的黏附依赖于α5β1整合素而非αvβ3整合素。抑制糖胺聚糖硫酸化或阻断成纤维细胞生长因子受体（FgfR）可降低B16V细胞系中的α5整合素。有趣的是，在敲低Ust的细胞中FgfR1的表达和激活降低。在体内，由于α5整合素的减少，B16VshUst细胞的肺转移受到抑制。作为概念验证，在人黑色素瘤细胞中敲低UST也显示ITGa5和黏附性降低。这是第一项表明Ust以及因此FgfR CS/DS低亲和力受体的2-O硫酸化会降低Itga5并导致黑色素瘤细胞黏附和迁移受损的研究。
