GnT-V参与调控整合素α5β1介导的单核细胞—内皮细胞的粘附和迁移
发布时间：2018-07-18 20:32
【摘要】：催化N-联接聚糖形成的糖基转移酶表达量的变化与多种细胞生物学行为有关，如：细胞粘附，细胞迁移，细胞增值及肿瘤细胞转移等。本研究中，我们观察到了N-乙酰氨基葡萄糖转移酶V(GnT-V)及其催化产物（β1，6-GlcNAc）表达量的变化与炎症反应有关：GnT-V参与调控整合素α5β1介导的单核细胞-内皮细胞的粘附和迁移。通过细胞粘附和迁移实验，我们观察到IFN-γ所处理的单核细胞THP-1对血管内皮细胞EA.hy926的粘附和侵袭能力明显增强；同时，QRT-PCR及westernblot方法检测到此类单核细胞内的GnT-V及其催化产物（β1,6-GlcNAc）表达量明显下降。采用GnT-V干扰质粒转染单核细胞THP-1后，其对血管内皮的粘附和迁移能力亦明显增强，与IFN-γ处理具有相同生物学效应。我们还发现，通过封闭整合素α5或β1亚基可以逆转GnT-V干扰及IFN-γ处理所增强的单核细胞粘附和侵袭能力；westernblot方法验证证明上述处理减少整合素α5或β1亚基表面β1,6-GlcNAc的糖链修饰，而并不影响单核细胞内整合素α5或β1亚基表达。在相关信号通路的研究中发现，GnT-V表达量的下降明显增强整合素介导的FAK磷酸化。同时，我们观察到FAK磷酸化增强可激活其下游信号通路-ERK，而ERK磷酸化抑制剂预处理IFN-γ刺激的单核细胞后其对内皮的黏附和侵袭能力无明显增强。综上，我们的研究第一次发现炎症因子处理单核细胞所诱导的炎症反应中，GnT-V活性明显下降，从而增强整合素α5β1介导的单核细胞-血管内皮黏附和迁移；同时推测整合素-FAK信号通路及FAK信号通路下游-ERK信号通路可能参与上述生物学效应的调控。GnT-V可能成为心脑血管炎症疾病研究及治疗的新靶点。
[Abstract]:The changes in the expression of glycosyltransferases catalyzing the formation of N-ligand are related to a variety of cell biological behaviors, such as cell adhesion, cell migration, cell proliferation and tumor cell metastasis. In this study, we observed that the expression of N-acetylglucosaminotransferase V (GnT-V) and its catalytic product (尾 _ 1N _ (6-GlcNAc) were related to the inflammatory response, and that: GnT-V was involved in regulating the adhesion and migration of integrin 伪 _ 5 尾 _ 1-mediated monocyte to endothelial cells. Through cell adhesion and migration experiments, we observed that the adhesion and invasion ability of THP-1 treated with IFN- 纬 on vascular endothelial cell EA.hy926 was significantly enhanced. At the same time, the expression of GnT-V and its catalytic product (尾 1N 6-GlcNAc) in these monocytes were detected by QRT-PCR and westernblot. After transfection with GnT-V interference plasmid, the adhesion and migration of THP-1 to vascular endothelium were significantly enhanced, which had the same biological effect as IFN- 纬 treatment. We also found that blocking integrin 伪 5 or 尾 1 subunit could reverse GnT-V interference and the enhanced adhesion and invasion ability of monocytes treated with IFN- 纬. Western blot showed that these treatments reduced the sugar chain modification of integrin 伪 5 or 尾 1 subunit surface 尾 1N 6-GlcNAc. The expression of integrin 伪 5 or 尾 1 subunit in monocytes was not affected. It was found that the decrease of GnT-V expression significantly enhanced integrin-mediated FAK phosphorylation. At the same time, we observed that enhanced phosphorylation of FAK activated its downstream signaling pathway -ERK, but the adhesion and invasion of IFN- 纬 stimulated monocytes were not significantly enhanced after pretreatment with ERK phosphorylation inhibitor. In conclusion, we found for the first time that the activity of GnT-V decreased significantly in the inflammatory response induced by inflammatory cytokines, which enhanced the adhesion and migration of integrin 伪 5 尾 1-mediated monocytes to vascular endothelium. It is also speculated that integrin-FAK signaling pathway and its downstream -ERK signaling pathway may be involved in the regulation of these biological effects. GnT-V may be a new target for the study and treatment of cardiovascular and cerebrovascular inflammatory diseases.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
本文编号：2132688
[Abstract]:The changes in the expression of glycosyltransferases catalyzing the formation of N-ligand are related to a variety of cell biological behaviors, such as cell adhesion, cell migration, cell proliferation and tumor cell metastasis. In this study, we observed that the expression of N-acetylglucosaminotransferase V (GnT-V) and its catalytic product (尾 _ 1N _ (6-GlcNAc) were related to the inflammatory response, and that: GnT-V was involved in regulating the adhesion and migration of integrin 伪 _ 5 尾 _ 1-mediated monocyte to endothelial cells. Through cell adhesion and migration experiments, we observed that the adhesion and invasion ability of THP-1 treated with IFN- 纬 on vascular endothelial cell EA.hy926 was significantly enhanced. At the same time, the expression of GnT-V and its catalytic product (尾 1N 6-GlcNAc) in these monocytes were detected by QRT-PCR and westernblot. After transfection with GnT-V interference plasmid, the adhesion and migration of THP-1 to vascular endothelium were significantly enhanced, which had the same biological effect as IFN- 纬 treatment. We also found that blocking integrin 伪 5 or 尾 1 subunit could reverse GnT-V interference and the enhanced adhesion and invasion ability of monocytes treated with IFN- 纬. Western blot showed that these treatments reduced the sugar chain modification of integrin 伪 5 or 尾 1 subunit surface 尾 1N 6-GlcNAc. The expression of integrin 伪 5 or 尾 1 subunit in monocytes was not affected. It was found that the decrease of GnT-V expression significantly enhanced integrin-mediated FAK phosphorylation. At the same time, we observed that enhanced phosphorylation of FAK activated its downstream signaling pathway -ERK, but the adhesion and invasion of IFN- 纬 stimulated monocytes were not significantly enhanced after pretreatment with ERK phosphorylation inhibitor. In conclusion, we found for the first time that the activity of GnT-V decreased significantly in the inflammatory response induced by inflammatory cytokines, which enhanced the adhesion and migration of integrin 伪 5 尾 1-mediated monocytes to vascular endothelium. It is also speculated that integrin-FAK signaling pathway and its downstream -ERK signaling pathway may be involved in the regulation of these biological effects. GnT-V may be a new target for the study and treatment of cardiovascular and cerebrovascular inflammatory diseases.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
【共引文献】
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2 李亚辉;沉默N-乙酰糖基转移酶V抑制CNE-2细胞的增殖和转移能力[D];南方医科大学;2011年
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