Zhao Yun-Peng, Xu Xin-Yun, Fang Meng, Wang Hao, You Qing, Yi Chang-Hong, Ji Jun, Gu Xing, Zhou Ping-Ting, Cheng Cheng, Gao Chun-Fang
Department of Laboratory Medicine, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China.
Department of General Surgery, Changzheng Hospital, Second Military Medical University, Shanghai, China.
PLoS One. 2014 Apr 14;9(4):e94536. doi: 10.1371/journal.pone.0094536. eCollection 2014.
The object of the study is to identify N-glycan profiling changes associated with gastric cancer and explore the impact of core-fucosylation on biological behaviors of human gastric cancer cells. A total of 244 subjects including gastric cancer, gastric ulcer and healthy control were recruited. N-glycan profiling from serum and total proteins in gastric tissues was analyzed by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis. The abundance of total core-fucosylated residues and the expression of enzymes involved in core-fucosylation were analyzed with lectin blot, quantitative reverse transcription-polymerase chain reaction, western blot, Immunohistochemical staining and lectin-histochemical staining. The recombinant plasmids of GDP-fucose transporter and α-1,6-fucosyltransferase (Fut8) were constructed and transfected into gastric cancer cell lines BGC-823 and SGC-7901. CCK-8 and wound healing assay were used to assess the functional impact of core-fucosylation modulation on cell proliferation and migration. Characteristic serum N-glycan profiles were found in gastric cancer. Compared with the healthy control, a trianntenary structure abundance, peak 9 (NA3Fb), was increased significantly in gastric cancer, while the total abundance of core-fucosylated residues (sumfuc) was decreased. Core-fucosylated structures, peak6(NA2F) and peak7(NA2FB) were deceased in gastric tumor tissues when compared with that in adjacent non-tumor tissues. Consistently, lens culinaris agglutinin (LCA)-binding proteins were decreased significantly in sera of gastric cancer, and protein level of Fut8 was decreased significantly in gastric tumor tissues compared with that in adjacent non-tumor tissues. Upregulation of GDP-Tr and Fut8 could inhibit proliferation, but had no significant influence on migration of BGC-823 and SGC-7901 cells. Core-fucosylation is down regulated in gastric cancer. Upregulation of core-fucosylation could inhibit proliferation of the human gastric cancer cells.
本研究的目的是鉴定与胃癌相关的N-聚糖谱变化，并探讨核心岩藻糖基化对人胃癌细胞生物学行为的影响。共招募了244名受试者，包括胃癌患者、胃溃疡患者和健康对照者。通过DNA测序仪辅助的荧光团辅助毛细管电泳分析血清和胃组织总蛋白中的N-聚糖谱。用凝集素印迹、定量逆转录-聚合酶链反应、蛋白质印迹、免疫组织化学染色和凝集素组织化学染色分析总核心岩藻糖基化残基的丰度和参与核心岩藻糖基化的酶的表达。构建GDP-岩藻糖转运体和α-1,6-岩藻糖基转移酶（Fut8）的重组质粒，并将其转染到胃癌细胞系BGC-823和SGC-7901中。采用CCK-8和伤口愈合试验评估核心岩藻糖基化调节对细胞增殖和迁移的功能影响。在胃癌患者中发现了特征性的血清N-聚糖谱。与健康对照相比，胃癌患者的三天线结构丰度（峰9，NA3Fb）显著增加，而核心岩藻糖基化残基的总丰度（sumfuc）降低。与相邻非肿瘤组织相比，胃癌组织中核心岩藻糖基化结构（峰6，NA2F和峰7，NA2FB）减少。同样，与相邻非肿瘤组织相比，胃癌患者血清中豆凝集素（LCA）结合蛋白显著减少，胃肿瘤组织中Fut8蛋白水平显著降低。GDP-Tr和Fut8的上调可抑制BGC-823和SGC-7901细胞的增殖，但对其迁移无显著影响。胃癌中核心岩藻糖基化下调。核心岩藻糖基化的上调可抑制人胃癌细胞的增殖。
