【摘要】 目的:探讨N-乙酰氨基葡萄糖基转移酶V(GnTV)表达受阻与内质网应激之间的关系。方法:应用RNAi技术,构建GnTVsiRNA表达质粒,抑制GnTV的表达,用RTPCR和Westernblot检测内质网应激中的重要分子GRP78(glucoseregulatedprotein,葡萄糖调节蛋白)、XBP1(Xboxbindingprotein1,X盒结合蛋白1)和PERK[RNAdependentproteinserine/threoninekinase(PKR)likekinase,RNA依赖蛋白丝/苏氨酸激酶的类似激酶]的表达变化。结果:GRP78的mRNA和蛋白质表达量升高;XBP1的mRNA发生剪接,蛋白分子量从33×103(33kD)变为54×103;PERK蛋白发生磷酸化。结论:GnTV受阻可导致内质网应激。更多还原

【Abstract】 Purpose:To study the connection between the blocking of N-acetylglucosaminyltransferase V(GnT-V) and endoplasmic reticulum stress(ER stress). Methods:The recombinant plasmid expressing siRNA of GnT-V was constructed and transfected into SMMC-7721. The key molecules such as GRP78,XBP1 and PERK during ER stress was analyzed in mRNA and protein levels. Results:GRP78 mRNA and protein levels were upregulated, the spliced forms of XBP1 mRNA and protein appeared and ER eIF2α kinase PERK was activated. Conclusions:These results suggest the blocking of GnT-V in SMMC-7721 induces cellular ER stress.更多还原