不同转移潜能膀胱癌细胞糖组相对定量分析

膀胱癌是发生在膀胱黏膜组织上的一种恶性肿瘤,是泌尿系统中最常见的恶性肿瘤,早期(非肌层浸润型膀胱癌)阶段的诊断和治疗是降低膀胱癌死亡率的最有效方式.肿瘤的发生过程与糖链表达的改变有着密切的关系,而定量分析膀胱癌发生过程中糖链的表达变化尚未有研究.本研究以2株人膀胱正常上皮细胞系(HCV29、HUCV1),1株非肌层浸润性膀胱癌细胞系(KK47),和3株浸润性膀胱癌细胞系(YTS1、J82、T24)为研究材料,应用本室建立的利用乙酰肼修饰糖链唾液酸,以及12C6]-和13C6]-苯胺同位素修饰糖链还原性末端技术,然后利用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS),进行膀胱上皮细胞不同病理状态的糖组相对定量分析.从6株细胞中共鉴定出52种N-连接糖链结构,并定量分析了不同类型的糖链在不同细胞中的分布差异,发现唾液酸化、岩藻糖化的N-连接糖链在膀胱癌肿瘤细胞恶化过程中呈现显著升高的趋势,同时平分型糖链和高甘露糖型N-连接糖链也呈表达升高趋势,说明这些糖链结构的表达变化与膀胱癌发生关系密切,从而有助于进一步阐明膀胱癌发生过程中糖链相关的分子机理.

Quantitative Analysis of N-Glycome of Different Bladder Cancer Cells

The bladder cancer, which is a kind of a malignant tumor in the bladder mucosa, is the most common malignancy tumor in the urinary system. Diagnosis at an early (nonmuscle-invasive) stage is the best way to reduce the mortality rate. Tumor malignancy is closely associated with alterations of glycan expression, but glycosylation status in bladder cancer has not been well studied. Four bladder cancer cell lines (KK47, YTS1, J82, T24) and two normal bladder mucosa cell line (HCV29, HUCV1) were used in our study. We developed a kind of labeling method to derivatize the sialic acid and derivatized the reducing terminal using ¹²C₆]- or ¹³C₆]-aniline, and the derivatized glycans were quantitatively analyzed by Matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF/TOF MS). Finally, 52 N-glycans of the bladder cancer were identified. And the sialic acid as well as the fucosylated in the bladder cancer were found significantly over-expressed. At the same time, the bisecting and high-mannose glycan were also high-expressed in the bladder cancer cells. This study provides some referential significance for the future study on the glycomics of bladder cancer.