糖基转移酶反义核酸对人脑胶质瘤细胞SWO-38作用的分子机理  

Molecular Mechanism of Antisense Glycosyltransferase Oligonucleotide Inhibiting Human Gliomas Cell Line SWO-38

作　　者：郑刚[1] 张积仁[1] 刘发全[1] 王雄文[1] 张新宇[2] 

机构地区：[1]第一军医大学珠江医院肿瘤科 [2]第一军医大学珠江医院骨科,广东广州510282

出　　处：《癌症》2002年第10期1095-1099,共5页Chinese Journal of Cancer

基　　金：国家自然科学基金(编号:39870813)

摘　　要：背景与目的:红细胞系列的鞘糖脂与很多人类肿瘤的生物学活性相关。α1,4半乳糖糖基转移(α1,4-galactosyltransferase,α1,4Gal-T)是红细胞系列鞘糖脂合成的特异的糖基转移酶。本实验目的是研究α1,4Ga反义寡核苷酸对脑胶质瘤细胞系SWO-38的作用及其分子机理。方法:人工合成α1,4Gal-T基因的反义寡核苷酸采用脂质体介导的基因转染方法将α1,4Gal-T基因反义核酸转入SWO-38细胞中。应用RT-PCR检测其对α4Gal-TmRNA表达的影响,应用克隆形成实验检测其对细胞增殖的作用,应用流式细胞术(FCM)和琼脂糖凝胶电检测细胞凋亡的发生,应用FCM检测细胞fas、p53、bax和bcl-2表达的变化。结果:细胞转染反义核酸后,经RT-P检测证实其α1,4Gal-TmRNA基因的表达下降;克隆形成实验显示细胞生长明显受到抑制(P<0.01);DNA电泳像呈凋亡表现;FCM检测表明,导入α1,4Gal-T基因反义核酸的SWO-38细胞凋亡明显增加(P<0.05),bcl-2蛋表达显著下降(P<0.01),而fas和bax蛋白表达显著提高(P<0.01),p53蛋白表达无显著变化。结论:α1,4Ga基因硫代反义核酸可诱导脑胶质瘤细胞系SWO-38细胞凋亡,其机理与fas、bcl-2和bax基因调控及其相互作用关。Background & Objective: It was reported that Globo series glycoshingolipids have relationship to many human cancers, and α1,4 galactosyltransferase(α1,4Gal T) oligonucleotide is the specific glycosyltransferase for the synthesis of Globo series glycoshingolipids.This study was designed to evaluate the effects of antisense α1,4Gal T oligonucleotide on human gliomas cell line SWO 38. Materials & Methods: SWO 38 glioma cells were reacted with 10 μmol/L antisense α1,4Gal T oligonucleotide for 72 hours by means of lipofectin transfection. The growth inhibition was detected by colony forming unit assay. DNA fragmentation was examined by flow cytometric analysis(FCM) and agarose gel eletrophoresis. The expressions of fas,p53,bax, and bcl 2 protein were determined by flow cytometric analysis (FCM). Results: Antisense α1, 4Gal T oligonucleotide signifantly inhibited cell growth (P< 0.01), induced the apoptosis (P< 0.05), downregulated expressoin of bcl 2 protein (P< 0.01) and upregulated expressions of fas and bax protein (P< 0.01), but did not influence p53 expression in glioma cell line SWO 38. Conclusions: Antisense α1,4Gal T oligonucleotide can significantly inhibit proliferation and induce apoptosis in human gliomas cell line SWO 38, which maybe due to the interactions of fas,bax, and bcl 2.

关 键 词：糖基转移酶 分子机理 脑胶质瘤 α1 4半乳糖糖基转移酶 反义核酸 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]