肝细胞癌细胞诱导和体外培养的大鼠肝星状细胞基因差异表达  

Analysis of differential gene expression in rat hepatic stellate cells activated by culture or hepatocellular carcinoma cell induction

作　　者：夏云红[1] 宋正己[2] 陈荣新[1] 叶胜龙[1] 孙瑞霞[1] 薛琼[1] 

机构地区：[1]复旦大学附属中山医院复旦大学肝癌研究所,上海200032 [2]复旦大学附属中山医院消化内科,上海200032

出　　处：《中华肿瘤杂志》2009年第3期164-169,共6页Chinese Journal of Oncology

基　　金：基金项目：国家重点基础研究（973）项目（2004CB518708）

摘　　要：目的从基因水平了解肝细胞癌（HCC）内的星状细胞（HSC）与正常肝HSC的异同。方法密度梯度离心分离HSC，用大鼠HCC细胞株C5F的条件培养基诱导HSC活化。cDNA微阵列比较静止、体外培养活化和诱导活化HSC之间22012个基因的表达，并通过实时逆转录聚合酶链反应（real-time RT-PCR）和Western blot检测进行验证。结果与静止HSC相比，体外培养HSC共有1672个基因差异表达，包括促炎症因子、细胞表面受体、黏附分子、信号转导分子、免疫因子等，其中1012个基因上调，660个基因下调。肿瘤诱导活化HSC有711个基因差异表达，其中420个基因上调，291个基因下调，有一部分基因表达与体外培养相同，部分基因如Raf1、Rac2、Adam17、Wnt6、MMP-9和TNF等，呈特异性表达。real-time RT-PCR和Western blot检测结果与cDNA微阵列检测结果相符。结论HCC细胞可特异性驱动HSC的活化，诱导活化的HSC在HCC细胞的浸润和转移中可能起重要作用。瘤内活化的HSC应作为研究HSC生物学功能的标准。Objective Hepatic stellate cells （HSC） in hepatocellular carcinoma （HCC） transdifferentiate into extraeellular matrix-producing myofibroblasts. Activated HSC can promote invasion and metastasis of HCC. To understand the differences of HSC in normal liver and HCC, we compared the gene expression patterns in HCC cell induction-activated and culture-activated rat HSC. Methods HSC were isolated by density centrifugation and exposed to conditioned medium from rat HCC cell line C5F. Expression of 22 012 genes in quiescent HSC, culture-activated HSC and HCC induction-activated HSC was analyzed by cDNA microarray and confirmed by real-time RT-PCR and Western blot. Results 1672 genes were differentially expressed in culture-activated HSC, including proinflammatory factors, cell adhesion molecules, cell surface receptors, signaling transduction molecules and immune factors. 711 genes were differentially expressed in HCC induction-activated HSC. Some of them were identical to those in culture-activated HSC. HCC Induction-activated HSC showed specific gene expression patterns, including Raf1, Rac2, Adam17, Wnt6, MMP-9 and TNF, suggesting that HCC cells can specifically induce HSC activation. Conclusion The gene expression patterns in HCC induction-activated HSC are different from those in culture-activated HSC. HCC induction-activated HSC may play a major role in the invasion and metastasis of HCC. In vivo activation should be considered as the standard for the study of HSC biology. HCC induction-activated HSC should be considered as the standard for HSC biology studies.

关 键 词：癌 肝细胞 星形细胞 基因表达 寡核苷酸序列分析 

分 类 号：R686[医药卫生—骨科学]