Retinoic acid-induced changes in polysialyltransferase mRNA expression and NCAM polysialylation in human neuroblastoma cells.

R. Seidenfaden, H. Hildebrandt
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引用次数: 15

Abstract

Polysialic acid (PSA) is a dynamically regulated carbohydrate modification of the neural cell adhesion molecule NCAM, which is implicated in neural differentiation and cellular plasticity. The cloning and characterization of two polysialyltransferases, termed ST8SiaII (STX) and ST8SiaIV (PST), opened up new perspectives in the search for factors that control this unique cell surface glycosylation. In vitro and transfection approaches revealed that ST8SiaII and ST8SiaIV are independently capable of synthesizing PSA on NCAM with slightly different specificities towards the major NCAM isoforms and glycosylation sites. Their overlapping but distinct expression patterns during brain development point towards an independent transcriptional regulation. However, the factors driving their joint or distinct expression, as well as the significance of divergent expression patterns in vivo, are not yet understood. In the present study, the mRNA expression of ST8SiaII and ST8SiaIV was comparatively analyzed in neuronal differentiation of PSA-positive human neuroblastoma cell lines induced by retinoic acid (RA), phorbolester, or growth factors. Using a semiquantitative RT-PCR strategy, we demonstrated a general decrease in the mRNA level of ST8SiaII upon differentiation of SH-SY5Y and LAN-5 cells. In contrast, a drastic increase of ST8SiaIV was specifically induced by RA-treatment of SH-SY5Y cells. To explore the significance of these changes, the cellular capacity to perform PSA synthesis and the degree of NCAM polysialylation were analyzed. Our data indicate that the increased expression of ST8SiaIV enables an accelerated polysialylation of NCAM, which, however, is not converted into higher amounts of PSA.
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维甲酸诱导的人神经母细胞瘤细胞中多唾液基转移酶mRNA表达和NCAM多唾液基化的变化。
聚唾液酸(PSA)是神经细胞黏附分子NCAM的一种动态调控碳水化合物修饰,与神经细胞分化和细胞可塑性有关。ST8SiaII (STX)和ST8SiaIV (PST)两种多唾液基转移酶的克隆和鉴定,为寻找控制这种独特细胞表面糖基化的因素开辟了新的视角。体外和转染方法表明,ST8SiaII和ST8SiaIV能够独立合成NCAM上的PSA,对NCAM主要亚型和糖基化位点的特异性略有不同。它们在大脑发育过程中重叠但不同的表达模式指向一个独立的转录调控。然而,驱动它们联合或不同表达的因素,以及不同表达模式在体内的意义尚不清楚。本研究比较分析了ST8SiaII和ST8SiaIV mRNA表达在视黄酸(RA)、磷酸酯(phorbolester)和生长因子诱导的psa阳性人神经母细胞瘤细胞系的神经元分化过程中。使用半定量RT-PCR策略,我们证明了SH-SY5Y和LAN-5细胞分化时ST8SiaII mRNA水平普遍下降。相反,ra处理SH-SY5Y细胞特异性诱导ST8SiaIV急剧增加。为了探讨这些变化的意义,我们分析了细胞进行PSA合成的能力和NCAM多唾液化的程度。我们的数据表明,ST8SiaIV表达的增加能够加速NCAM的多唾液化,然而,它不会转化为更高量的PSA。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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