Abstract

Bladder cancer is the ninth most frequently diagnosed cancer worldwide, and there is a need to develop new biomarkers for staging and prognosis of this disease. Here we report that cell lines derived from low-grade and high-grade bladder cancers exhibit major differences in expression of glycans in surface glycoproteins. We analyzed protein glycosylation in three low-grade bladder cancer cell lines RT4 (grade-1-2), 5637 (grade-2), and SW780 (grade-1), and three high-grade bladder cancer cell lines J82COT (grade-3), T24 (grade-3) and TCCSUP (grade-4), with primary bladder epithelial cells, A/T/N, serving as a normal bladder cell control. Using a variety of approaches including flow cytometry, immunofluorescence, glycomics and gene expression analysis, we observed that the low-grade bladder cancer cell lines RT4, 5637 and SW780 express high levels of the fucosylated Lewis-X antigen (Lex, CD15) (Galβ1–4(Fucα1–3)GlcNAcβ1-R), while normal bladder epithelial A/T/N cells lack Le^x expression. T24 and TCCSUP cells also lack Le^x, whereas J82COT cells express low levels of Le^x. Glycomics analyses revealed other major differences in fucosylation and sialylation of N-glycans between these cell types. O-glycans are highly differentiated, as RT4 cells synthesize core 2-based O-glycans that are lacking in the T24 cells. These differences in glycan expression correlated with differences in RNA expression levels of their cognate glycosyltransferases, including α1–3/4-fucosyltransferase genes. These major differences in glycan structures and gene expression profiles between low- and high-grade bladder cancer cells suggest that glycans and glycosyltransferases are candidate biomarkers for grading bladder cancers.

中文翻译：

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低级别与高级别膀胱癌细胞系中糖基化和岩藻糖基转移酶表达的主要差异

 抽象的

膀胱癌是全球第九大最常诊断的癌症，需要开发新的生物标志物用于该疾病的分期和预后。在这里，我们报告来自低级别和高级别膀胱癌的细胞系在表面糖蛋白中聚糖的表达方面表现出主要差异。我们分析了三种低级别膀胱癌细胞系 RT4（1-2 级）、5637（2 级）和 SW780（1 级）以及三种高级别膀胱癌细胞系 J82COT（1-2 级）中的蛋白质糖基化。 3)、T24（3 级）和 TCCSUP（4 级），具有原代膀胱上皮细胞、A/T/N，作为正常膀胱细胞对照。使用流式细胞术、免疫荧光、糖组学和基因表达分析等多种方法，我们观察到低级别膀胱癌细胞系 RT4、5637 和 SW780 表达高水平的岩藻糖基化 Lewis-X 抗原（Lex、CD15）（Galβ1） –4(Fucα1–3)GlcNAcβ1-R)，而正常膀胱上皮A/T/N细胞缺乏Le ^x表达。 T24 和 TCCSUP 细胞也缺乏 Le ^x ，而 J82COT 细胞表达低水平的 Le ^x 。糖组学分析揭示了这些细胞类型之间N-聚糖的岩藻糖基化和唾液酸化的其他主要差异。 O-聚糖是高度分化的，因为 RT4 细胞合成 T24 细胞所缺乏的基于核心 2 的O-聚糖。这些聚糖表达的差异与其同源糖基转移酶（包括 α1-3/4-岩藻糖基转移酶基因）的 RNA 表达水平差异相关。 低级别和高级别膀胱癌细胞之间聚糖结构和基因表达谱的这些主要差异表明聚糖和糖基转移酶是膀胱癌分级的候选生物标志物。