人肝癌Bel-7402细胞及Bel-7402/5-FU细胞的N-连接聚糖的差异分析

Differential Analysis of N-glycans from Human Hepatocellular Carcinoma Bel-7402 and Bel-7402/5-FU Cells

以培养的人肝癌细胞Bel-7402及其5-氟尿嘧啶(5-FU)耐药细胞(Bel-7402/5-FU)为研究对象,通过使用快速PNGase F酶切,结合TMPP-Ac-OSu和甲胺化共衍生方法,对两者的总蛋白和分泌蛋白的N-连接聚糖进行了基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)分析。从两种细胞总蛋白中共鉴定到56种N-连接聚糖,分泌蛋白中鉴定到38种N-连接聚糖。5-FU与Bel-7402相比,耐药细胞岩藻糖化唾液酸糖型在总蛋白和分泌蛋白中都显著升高;高甘露糖型N-聚糖在总蛋白中下降,而在分泌蛋白中显著增加。本研究为进一步探索肿瘤耐药提前诊断提供糖链标志物起到一定的参考作用,并为后续解决癌症治疗的耐药提供理论依据。

In this study,an optimized co-derivatization method was successfully used to reveal the difference in cellular N-glycan profiling of Bel-7402 and Bel-7402/5-FU.56 and 38 N-glycans were identified in total proteins and secreted proteins of these two cell lines,respectively.The results showed that core-fucosylated complex glycans in Bel-7402/5-FU cells were significantly increased when it was compared with Bel-7402 cells.In addition,the high mannose type N-glycan decreased in total protein and increased in secretory protein in Bel-7402/5-FU cells.This study plays a certain reference role in providing glycan biomarkers for the further exploration of advanced diagnosis of cancer drug resistance,and provides a theoretical basis for subsequent solution of drug resistance in cancer treatment.