糖基转移酶C3ORF39的亚细胞内定位及组织分布特征研究  

Subcellular location of glycosyltransferase C3ORF39 in HepG2 cells, and distribution in different tissues

作　　者：魏红山[1,2] 李学永[3] 肖凡[1,2] 郝晓花[1,2] 黄玉波[1,2] 杜雅菊[4] 

机构地区：[1]首都医科大学附属北京地坛医院传染病研究所 [2]新发突发传染病研究北京市重点实验室,北京100015 [3]河北省邯郸市中心医院急诊科 [4]哈尔滨医科大学第二附属医院消化科

出　　处：《中华实验和临床感染病杂志（电子版）》2013年第4期4-6,共3页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学基金(No.81071411;81271901)

摘　　要：目的 Notch分子是一个高度糖基化修饰的信号分子,但催化其O-葡聚糖中木糖转糖基反应的糖基转移酶至今未能明确。本研究旨在克隆表达人类糖基转移酶61家族中的C3orf39,初步明确其细胞内定为及部分组织分布特征。方法体外克隆表达C3ORF39重组蛋白,免疫新西兰兔制备该重组蛋白的多克隆抗体;采用免疫组织化学方法明确该基因编码的糖基转移酶在部分组织细胞中的分布特征。构建荧光重组蛋白编码质粒,转染HepG2细胞,利用激光共聚焦显微镜观察该糖基转移酶可能的亚细胞定位。结果制备的C3ORF39重组蛋白多克隆抗体具有较高的免疫活性和特异性。免疫组织化学分析显示,该糖基因主要表达与肝细胞、肠上皮细胞、肾小管上皮细胞和胃腺体上皮细胞;部分表达于淋巴结、脾脏、间质细胞。激光共聚焦观察显示,该糖基转移酶定位于内质网-Golgi体分泌途径上游。结论糖基因C3orf39定位于分泌途径中的内质网或cis-Golgi体部分,广泛表达与上皮组织和部分间质细胞。Objective Notch molecular is a highly glycosylated signal protein, but the glycosyltransferase catalyzing Xylose transferase to O-glycan is remain elucidated. The present study was designed to prepare C3ORF39 recombinant protein and its polycolonal antibody, and to determine its subcellular location and tissue distribution. Methods The C3ORF39 recombinant protein was prepared in vitro, and immunized rabbit to prepare polyclonal antibody. The tissue distribution of C3ORF39 was examined in different tissue by immunohistochemistry method. The subcellular location was determined by laser confocal fluorescence microscopy. Results The prepared polyclonal antibody showed higher activity and speciifcity. The immunohistochemical staining demonstrated that C3ORF39 mainly scattered in hepatocytes, intestinal epithelial cells and renal tubular epithelial cells, gastric epithelial cells;moderately expressed in interstitial cells of lymph node and spleen. C3ORF39 was located in endoplasmic reticulum and cis-Golgi complexity of HepG2 cells. Conclusions The C3ORF39 was located in proximity of secretory pathway of HepG2 cells, and mainly scattered in epithelial cells and moderately expressed in some interstitial cells.

关 键 词：糖基因 C3ORF39 糖基转移酶 NOTCH信号途径 肝细胞癌 

分 类 号：R735.7[医药卫生—肿瘤]