抑制糖基转移酶GnT-V的表达可增强胶质瘤细胞放射敏感性

Inhibition of Glycosyltransferase GnT-V Expression Enhances the Radiosensitivity of Glioma Cells

目的:探讨N-乙酰氨基葡萄糖转移酶V(GnT-V)介导的N-糖基化修饰在胶质瘤细胞放射敏感性中的作用及调控机制。方法:采用RT-PCR和Western blot技术筛选GnT-V高表达的胶质瘤细胞;运用RNA干扰技术抑制GnT-V基因和蛋白表达,流式细胞术和荧光显微镜观察细胞N-糖基化修饰改变;4 Gy X-射线处理后分析细胞放射敏感性差异。结果:成功筛选GnT-V高表达的U251细胞;下调U251细胞中GnT-V的表达,可阻断N-糖链合成,逆转X-射线诱导的G2/M期阻滞,促进细胞凋亡。结论:抑制GnT-V的表达可通过阻断N-糖基化修饰增强胶质瘤细胞的放射敏感性。

Objective To investigate the effects of N-acetylglucosaminyltransferase V（ GnT-V）-mediated N-glycosylation on the radiosensitivity of human glioma cells and its possible mechanism. Methods The GnT-V over-expressing glioma cell line was selected by RT-PCR and Western blot. RNA interference was used to inhibit the expression of GnT-V mRNA and protein,and the changes of N-glycosylation were detected by flow cytometry and fluorescence microscopy. Cell radiosensitivity differences were analyzed after 4 Gy X-ray radiation. Results U251 cells with high GnT-V expression were successfully selected. Down-regulation of GnT-V in U251 cells could block the synthesis of N-glycans,reverse X-ray induced G2/M arrest and promote apoptosis. Conclusion The inhibition of GnT-V expression enhances the radiosensitivity of glioma cells by blocking N-glycosylation.