目的:通过siRNA干扰技术研究FUT6基因沉默对Hep G2细胞侵袭和迁移能力的影响。方法:靶向FUT6的siRNA转染肝癌细胞株Hep G2后,通过western blot检测FUT6蛋白的表达,通过流式细胞术检测肝癌细胞表面抗原唾液酸路易斯糖(Sialyl Lewis X,s L...目的:通过siRNA干扰技术研究FUT6基因沉默对Hep G2细胞侵袭和迁移能力的影响。方法:靶向FUT6的siRNA转染肝癌细胞株Hep G2后,通过western blot检测FUT6蛋白的表达,通过流式细胞术检测肝癌细胞表面抗原唾液酸路易斯糖(Sialyl Lewis X,s Le X)表达情况。Transwell小室方法进行细胞迁移和侵袭实验检测肝癌细胞株Hep G2转移和侵袭能力的变化。结果:实验分为3组,分别命名为control(空白对照组),mock(阴性对照scrambled-siRNA)和siRNA-FUT6(实验组)。实验组FUT6蛋白及s Le X抗原较空白对照组明显降低,实验组细胞的迁移能力及侵袭能力下降。结论:siRNA靶向沉默FUT6减少了FUT6蛋白的表达,减少了肿瘤相关抗原s Le X的表达,削弱Hep G2细胞在体外的迁移和侵袭能力,FUT6可能成为肝癌基因治疗的新靶点。展开更多
E-selectin is a cell-adhesion molecule of the vascular endothelium that promotes essential leukocyte rolling in the early inflammatory response by binding to glycoproteins containing the tetrasaccharide sialyl Lewisx(...E-selectin is a cell-adhesion molecule of the vascular endothelium that promotes essential leukocyte rolling in the early inflammatory response by binding to glycoproteins containing the tetrasaccharide sialyl Lewisx(sLex).Efficient leukocyte recruitment under vascular flow conditions depends on an increased lifetime of E-selectin/ligand complexes under tensile force in a so-called catch-bond binding mode.Co-crystal structures of a representative fragment of the extracellular E-selectin region with sLex and a glycomimetic antagonist thereof reveal an extended E-selectin conformation,which is identified as a high-affinity binding state of E-selectin by molecular dynamics simulations.Small-angle X-ray scattering experiments demonstrate a direct link between ligand binding and E-selectin conformational transition under static conditions in solution.This permits tracing a series of concerted structural changes connecting ligand binding to conformational stretching as the structural basis of E-selectin catch-bond-mediated leukocyte recruitment.The detailed molecular view of the binding site paves the way for the design of a new generation of selectin antagonists.This is of special interest,since their therapeutic potentialwas recently demonstrated with the pan-selectin antagonists GMI-1070(Rivipansel).展开更多
文摘目的:通过siRNA干扰技术研究FUT6基因沉默对Hep G2细胞侵袭和迁移能力的影响。方法:靶向FUT6的siRNA转染肝癌细胞株Hep G2后,通过western blot检测FUT6蛋白的表达,通过流式细胞术检测肝癌细胞表面抗原唾液酸路易斯糖(Sialyl Lewis X,s Le X)表达情况。Transwell小室方法进行细胞迁移和侵袭实验检测肝癌细胞株Hep G2转移和侵袭能力的变化。结果:实验分为3组,分别命名为control(空白对照组),mock(阴性对照scrambled-siRNA)和siRNA-FUT6(实验组)。实验组FUT6蛋白及s Le X抗原较空白对照组明显降低,实验组细胞的迁移能力及侵袭能力下降。结论:siRNA靶向沉默FUT6减少了FUT6蛋白的表达,减少了肿瘤相关抗原s Le X的表达,削弱Hep G2细胞在体外的迁移和侵袭能力,FUT6可能成为肝癌基因治疗的新靶点。
基金supported by the Swiss National Science Foundation(grant number 200020_129935 and R’EQUIP 145023).
文摘E-selectin is a cell-adhesion molecule of the vascular endothelium that promotes essential leukocyte rolling in the early inflammatory response by binding to glycoproteins containing the tetrasaccharide sialyl Lewisx(sLex).Efficient leukocyte recruitment under vascular flow conditions depends on an increased lifetime of E-selectin/ligand complexes under tensile force in a so-called catch-bond binding mode.Co-crystal structures of a representative fragment of the extracellular E-selectin region with sLex and a glycomimetic antagonist thereof reveal an extended E-selectin conformation,which is identified as a high-affinity binding state of E-selectin by molecular dynamics simulations.Small-angle X-ray scattering experiments demonstrate a direct link between ligand binding and E-selectin conformational transition under static conditions in solution.This permits tracing a series of concerted structural changes connecting ligand binding to conformational stretching as the structural basis of E-selectin catch-bond-mediated leukocyte recruitment.The detailed molecular view of the binding site paves the way for the design of a new generation of selectin antagonists.This is of special interest,since their therapeutic potentialwas recently demonstrated with the pan-selectin antagonists GMI-1070(Rivipansel).
