大鼠肝卵圆样细胞恶性转化过程中的基因表达谱分析  

Analysis of gene expression profile during malignant transformation of rat fiver oval-like cells

作　　者：李雪飞[1] 李岩[1] 高东梅[1] 康晓楠[2] 郭坤[1] 王念[2] 赵燕[1] 孙璐[2] 刘银坤[1] 

机构地区：[1]复旦大学附属中山医院肝癌研究所,上海200032 [2]复旦大学生物医学研究院癌症中心

出　　处：《中华肝脏病杂志》2008年第10期767-771,共5页Chinese Journal of Hepatology

基　　金：国家“863”项目（2006AA02A308）

摘　　要：目的分析肝卵圆细胞恶性转化过程中基因表达谱的改变，探讨差异基因的意义。方法经直接致癌剂N-甲基-N＇-硝基N-亚硝基胍启动的大鼠肝卵圆细胞样WBF344暴露于过氧化氢，每周1次，重复21次诱发其恶性转化。通过形态学、遗传学和软琼脂克隆形成等实验验证细胞的转化。并应用四甲基偶氮唑盐法检测转化细胞的增殖能力。采用大鼠癌通路发现者功能芯片比较恶性转化过程中正常对照（WB），WB-5（过氧化氢刺激5次），WB21（过氧化氢刺激21次）3组细胞基因表达的差异。结果转化的细胞获得了异倍体核型，具有非停泊依赖生长特性。电镜下转化细胞的细胞器增多，出现缝隙连接和微绒毛等结构。基因表达谱分析显示共有21个基因差异表达，主要参与调节细胞增殖、凋亡、细胞黏附运动等，其中连续上调的有pten，连续下调的有cdknla，而先上调再下调的基因有myc、fos、casp8等。结论细胞增殖和凋亡的异常在卵圆细胞恶性转化过程中起重要作用，而cdknla、pten、myc、los等基因可能是起关键作用的分子。Objectives To investigate the changes of gene expression profile during malignant transformation of rat liver oval-like cells and to analyze the significances of these changes. Methods MNNG initiated WB-F344 cells were exposed to H202 once a week （repeated 21 times） to induce their malignant transformation. The characteristics of the transformed cells were confuxned by morphology, genetics and soft agar assay. Then, gene expression profiles of the transformed cells at different time points were evaluated using rat Cancer PathwayFinder Oligo Microarray. Results The transformed cells possessed heteroploid karyotypes with anchoring-independent growth characteristics. Transmission electron microscopy showed that the transformed cells had more cellular organelles, gap junctions and microvilli than the controls. The 21 differential expression genes were mainly involved in regulatiing cell proliferation, apoptosis, adhesion and motility. The expression of PTEN was gradually up-regulated and cdknl ct was gradually down-regulated in three groups of cells. Myc, fos, and casp-8 were first up-regulated in WB-5 cells and then down-regulated in WB-21 cells. Conclusion The disorder of proliferation and apoptosis may play an important role in malig- nant transformation of WB cells. Cdknl a, PTEN, myc and fos may be crucial factors involved in the process.

关 键 词：癌 肝细胞 细胞转化 肿瘤 细胞凋亡 肝卵圆细胞 

分 类 号：R735.7[医药卫生—肿瘤]