中华实验和临床感染病杂志（电子版）
中華實驗和臨床感染病雜誌（電子版）
중화실험화림상감염병잡지（전자판）
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL INFECTIOUS DISEASES(ELECTRONIC VERSION)
2013年
4期
487-490
,共4页
魏红山%李学永%肖凡%郝晓花%黄玉波%杜雅菊
魏紅山%李學永%肖凡%郝曉花%黃玉波%杜雅菊
위홍산%리학영%초범%학효화%황옥파%두아국
糖基因%C3ORF39%糖基转移酶%Notch信号途径%肝细胞癌
糖基因%C3ORF39%糖基轉移酶%Notch信號途徑%肝細胞癌
당기인%C3ORF39%당기전이매%Notch신호도경%간세포암
Glycogene%C3ORF39%Glycosyltransferase%Notch signal pathway%Hepatocellular carcinoma
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。
目的： Notch分子是一箇高度糖基化脩飾的信號分子，但催化其O-葡聚糖中木糖轉糖基反應的糖基轉移酶至今未能明確。本研究旨在剋隆錶達人類糖基轉移酶61傢族中的C3orf39，初步明確其細胞內定為及部分組織分佈特徵。方法體外剋隆錶達C3ORF39重組蛋白，免疫新西蘭兔製備該重組蛋白的多剋隆抗體；採用免疫組織化學方法明確該基因編碼的糖基轉移酶在部分組織細胞中的分佈特徵。構建熒光重組蛋白編碼質粒，轉染HepG2細胞，利用激光共聚焦顯微鏡觀察該糖基轉移酶可能的亞細胞定位。結果製備的C3ORF39重組蛋白多剋隆抗體具有較高的免疫活性和特異性。免疫組織化學分析顯示，該糖基因主要錶達與肝細胞、腸上皮細胞、腎小管上皮細胞和胃腺體上皮細胞；部分錶達于淋巴結、脾髒、間質細胞。激光共聚焦觀察顯示，該糖基轉移酶定位于內質網-Golgi體分泌途徑上遊。結論糖基因C3orf39定位于分泌途徑中的內質網或cis-Golgi體部分，廣汎錶達與上皮組織和部分間質細胞。
목적： Notch분자시일개고도당기화수식적신호분자，단최화기O-포취당중목당전당기반응적당기전이매지금미능명학。본연구지재극륭표체인류당기전이매61가족중적C3orf39，초보명학기세포내정위급부분조직분포특정。방법체외극륭표체C3ORF39중조단백，면역신서란토제비해중조단백적다극륭항체；채용면역조직화학방법명학해기인편마적당기전이매재부분조직세포중적분포특정。구건형광중조단백편마질립，전염HepG2세포，이용격광공취초현미경관찰해당기전이매가능적아세포정위。결과제비적C3ORF39중조단백다극륭항체구유교고적면역활성화특이성。면역조직화학분석현시，해당기인주요표체여간세포、장상피세포、신소관상피세포화위선체상피세포；부분표체우림파결、비장、간질세포。격광공취초관찰현시，해당기전이매정위우내질망-Golgi체분비도경상유。결론당기인C3orf39정위우분비도경중적내질망혹cis-Golgi체부분，엄범표체여상피조직화부분간질세포。
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.
