肝癌细胞N-乙酰氨基葡萄糖基转移酶V表达受阻与内质网应激之间的关系  

Relationship between blocking of N-acetylglucosaminyltransferase V in hepatocarcinoma cells and cellular endoplasmic reticulum stress

作　　者：徐莺莺[1] 陆祎[1] 张毅[1] 申宗侯[1] 

机构地区：[1]复旦大学上海医学院生物化学与分子生物学系,上海200032

出　　处：《中国癌症杂志》2005年第2期130-134,共5页China Oncology

基　　金：国家自然科学基金项目(No.30370343)。

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。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.

关 键 词：内质网应激 N-乙酰氨基葡萄糖基转移酶V RNAI 

分 类 号：R735.7[医药卫生—肿瘤]