反义MUC2体外抑制胃癌细胞的增殖活性  

MUC2 antisense oligodeoxynucleotide inhibits proliferation of human gastric carcinoma cells in vitro

作　　者：张晓艳[1] 易永芬[1] 肖春卫[2] 

机构地区：[1]重庆医科大学病理教研室,重庆市400016 [2]深圳市西乡人民医院病理科,广东省深圳市518102

出　　处：《世界华人消化杂志》2005年第11期1278-1282,共5页World Chinese Journal of Digestology

基　　金：重庆市卫生局科研基金资助项目(渝卫科教);No.2000-48~~

摘　　要：目的:研究反义核酸技术体外抑制MUC2的表达,观察MUC2反义脱氧寡核苷酸(antisenseoligodeoxvnucleotide,ASODN)对胃癌细胞生长的抑制作用.方法:应用硫代磷酸修饰的MUC2ASODN经阳离子脂质体包裹后转染入人胃癌细胞株SGC7901,采用MTT法,形态学观察,免疫组化法及流式细胞仪检测MUC2ASODN对胃癌细胞的增殖抑制作用.结果:不同浓度ASODN均能抑制SGC7901细胞的增殖,在48h时最强,0.5μmol/LASODN对细胞的抑制率为55%,随时间延长抑制作用逐渐减弱.SGC7901细胞转染MUC2ASODN后,与对照组相比,光镜下观察到细胞数量减少,体积变小,核分裂明显减少,同时可见较多的坏死.流式细胞仪检测发现细胞主要被阻滞在S期.透射电镜下见细胞线粒体肿胀,细胞内脂滴增多,髓样结构,染色质边集等.免疫组化S-P法染色显示转染ASODN后,SGC7901细胞中MUC2,nm23蛋白表达水平明显降低,p16蛋白表达明显增强.结论:转染MUC2反义寡核苷酸能有效抑制胃癌细胞株SGC7901的增殖.AIM: To investigate the inhibitory effect of mucin gene (MUC2) antisense oligodeoxynucleotides (ASODNs) on proliferation of gastric cancer cells SGC7901. METHODS: Phosphorothioate MUC2 ASODNs were synthesized and then transfected into gastric carcinoma SGC7901 cells mediated by lipofectin. The inhibitory effects on cell proliferation were determined by MTT method, light and electronic microscopy, flow cytometry (FCM) and immunohistochemistry. RESULTS: MUC2 ASODN at varied concentration significantly inhibited the growth of SGC7901 cells in a dose-and time-dependent manner. The inhibitory peak appeared at 48th hour after transfection, and the inhibition rate reached 55% when the concentration of MUC2 ASODN was 0.5 nmol/L. After transfected with MUC2 ASODN, SGC7901 cells decreased in number, volume and karyokinesis, increased in necrosis under light microscopy. And most of the cells arrested in S phase. Swollen mitochondrion, increased liposomes, myelin figures, chromatin margination were found under electronic microscopy. Immunohis-tochemistry indicated that after transfected with MUC2 ASODN, the expression of MUC2 and nm23 protein were down-regulated, but the expression of p16 protein was up-regulated. CONCLUSION: MUC2 ASODN transfection can significantly inhibit the proliferation of gastric carcinoma SGC7901 cells.

关 键 词：MUC2 体外抑制 增殖活性 SGC7901细胞 流式细胞仪检测 ASODN 反义脱氧寡核苷酸 p16蛋白表达 反义核酸技术 胃癌细胞生长 人胃癌细胞株 增殖抑制作用 S-P法染色 蛋白表达水平 反义寡核苷酸 脂质体包裹 形态学观察 免疫组化法 

分 类 号：R735.2[医药卫生—肿瘤] R285.5[医药卫生—临床医学]