姑息性肝切除术后残癌转移潜能及其基因功能网络变化的实验观察  

Experimental observation on the alteration of metastatic potential and its gene function net of residual hepatocellular carcinoma following palliative liver resection

作　　者：黄修燕[1] 黄自丽[2] 汤钊猷[3] 徐彬[4] 郑起[1] 周俭[3] 王鲁[3] 叶胜龙[3] 

机构地区：[1]上海交通大学附属第六人民医院普外科,200233 [2]皖南医学院附属弋矶山医院神经 [3]复旦大学肝癌研究所 [4]同济大学附属第十人民医院普外科

出　　处：《中华医学杂志》2010年第18期1278-1282,共5页National Medical Journal of China

基　　金：国家重点基础研究发展计划（973）项目（2004CB518708）

摘　　要：目的 观察姑息性肝切除对肝脏残癌转移潜能的影响及其转移相关基因功能网络的变化.方法 以高转移人肝癌细胞（MHCC97H）建立裸鼠原位移植瘤模型,14 d后将36只成模裸鼠随机分成3组：姑息性肝切除组（获取肝癌标本A、B）、对照组1（假手术组,获取肝癌标本C1）、对照组2（无干预组,获取肝癌标本C2）.姑息术后14 d每组随机处死6只裸鼠,采用肿瘤转移相关芯片检测肝癌基因表达,使用综合型GEArray分析配套软件分析芯片数据.差异基因显著性分析、分组关联度分析及支持向量机用于筛选肿瘤转移相关标志基因;应用基于特征性致病基因的基因聚类和多维量表构建基因功能网络,比较网络密度、凝聚度以及枢纽基因的变化特点;Real-time PCR用于检测mRNA表达.姑息术后35 d处死剩余裸鼠,检测肺转移.SAS 8.2软件包用于统计分析.结果 组间存在显著性差异基因表达,姑息组基因网络密度（B,0.0670）较对照组（A,0.0145;C1,0.0210;C2,0.0146）上调;残癌基因网络凝聚度（B,0.1940）明显高于自身对照组（A,0.0098）.编码转移消失蛋白B（MIM-B）的人源性转移抑制基因1（MTSS1）处于残癌基因功能网络核心位置,残癌MIM-B mRNA相对表达水平（B,0.283±0.023）较对照组上调（A,0.142±0.018;C1,0.177±0.054;C2,0.156±0.017,P＜0.05）.术后35 d,姑息组肺转移结节数目（14.3±4.7）多于假手术组（8.7±3.6,P＜0.01）.结论 姑息性肝切除增强残癌肺转移潜能,MIM-B蛋白可能是肝癌患者重要的干预靶点之一.Objective To investigate the effects of palliative liver resection on metastatic potential and its gene function net of residual hepatocellular carcinoma （HCC） in nude mice model. Methods Orthotopic HCC model was established by implantation of human HCC cell line MHCC97H xenografts with a high metastatic potential. Thirty-six HCC-bearing nude mice were randomized into 3 groups at 14 days postoperation, including palliative resection group （ HCC samples A, B）, control group 1 （ sham operation,HCC sample C1 ） and control group 2 （without intervention, HCC sample C2）. Six mice in each group were sacrificed by cervical dislocation at 14 days after palliative resection. Oligo tumor metastasis microarray and GEArray expression analysis suite software were employed for gene analysis. The methods of support vector machine （SVM）, gene significance analysis and gene correlation degree analysis were used to search the markers capable of differentiating the metastatic potential of HCC. Gene function net was constructed based on special gene clustering analysis and multi-dimensional scale. Real-time PCR was applied to detect the mRNA expression. The remaining mice were sacrificed at 35 days after palliative resection for the examination of pulmonary metastasis. SAS 8.2 software was used for statistical analysis. Results Gene analysis showed that for different gene expression among groups, the density of gene net in palliative group （B, 0.0670） were higher than those in control groups （A, 0.0145; C1, 0.0210; C2, 0.0146）, the condensation degree of gene net in residual HCC （ B, 0. 1940） was higher than that in its own control group （A, 0. 0098 ）. Human gene metastasis suppressor 1 （MTSS1） that encodes a protein of missing in metastasis B （MIM-B） was situated in the central position of gene function net of residual HCC. The relative MIM-B mRNA expression examined by Real-time PCR was significantly up-regulated in residual HCC （B,0.283 ±0.023） as comparison was made among groups �

关 键 词：癌 肝细胞 姑息疗法 肿瘤转移 基因表达 裸鼠模型 

分 类 号：R735.7[医药卫生—肿瘤]