ID-1在肝癌发生和发展中作用分析进展.pdf

SiRNA-ID-1 对人肝癌细胞HepG2 裸鼠皮下移植瘤生长的影响 研究生：施纯礼 导师：朱月永副教授 中文摘要 目的：观察以RNA 干扰技术沉默ID-1 表达对人肝癌细胞HepG2 裸鼠皮下移植 瘤生长的抑制作用，以及对移植瘤组织bcl-2、Ki-67 、PCNA 和VEGF 表达的影 响，探讨抑制ID-1 表达对肝癌生长抑制的可能机制。 方法：向BALB/C-nu 系雄性裸鼠皮下注射HepG2 细胞建立裸鼠荷瘤模型，成瘤 后随机将裸鼠分为空白组、转染对照组和 siRNA-ID-1 处理组，在第 1、4 、7、 10 和13d 分别注射等量的生理盐水、control-siRNA 和甲基化的siRNA-ID-1 。每 周测量肿瘤体积，绘制肿瘤生长曲线。于第 28d 处死裸鼠，取肿瘤组织行常规 HE 染色，RT-PCR 和免疫组织化学(SP 法)分别检测肿瘤组织中ID-1、bcl-2、Ki-67 、 PCNA 及VEGF 的mRNA 和蛋白表达。 结果：1、siRNA-ID-1 处理组裸鼠肿瘤体积较空白组和转染对照组的体积小，差 别具有统计学意义(P0.05) ；HE 染色结果显示转染siRNA-ID-1 后肝癌细胞凋亡 明显增多，空白组及转染对照组瘤细胞排列密集，细胞界限不清，生长活跃，核 大深染，有较多核分裂象。2 、siRNA-ID-1 处理组肝癌组织中ID-1、bcl-2、Ki-67 、 PCNA 及VEGF mRNA 及蛋白表达均明显降低，差别具有统计学意义(P0.05) 。 结论：以siRNA 技术沉默ID-1 表达可通过抑制细胞增殖、诱导细胞凋亡而抑制 肿瘤的生长，提示ID-1 在抗肿瘤治疗中的潜在作用。 关键词：siRNA ID-1 肝癌 移植瘤 细胞增殖 2 The effects of RNA interference targeting ID-1 on the growth of hepatocellular carcinoma xenografts in nude mice Postgraduate shichunli Tutor Prof. zhuyueyong ABSTRACT AIMS ：To investigate the inhibition effects of RNA interference targeting ID-1(si-RNA-ID-1) on the growth of Hepatocellular Carcinoma(HCC) in nude mice and the potential mechanism of ID-1 in the developemnt of HCC. METHODS ：HCC xenografts were developed by injecting HepG2 cells into BALB/C-nu nude mice.The tumor-bearing mice were randomly divided into 3 groups, including blank control group,control-siRNA group and siRNA-ID-1 group.Each group was treated by isovolumetric normal saline,control-siRNA or siRNA-ID-1 on day 1,4,7,10 and 13, respectively. The growth curves of HCC xenografts were made by the tumor sizes, which were measured per week.. The nude mice were executed on day 28. The tumor tissue were prepared for the HE