摘要
目的构建肝细胞癌（肝癌）转移相关分子模型并筛选关键分子，用于评估肝癌转移潜能。方法以高转移人肝癌细胞MHCC97H建立裸鼠原位移植瘤模型，14d后将36只成模裸鼠随机分成姑息性肝切除组（获取肝癌标本A、B）、假手术组（获取肝癌标本C1）和对照组（获取肝癌标本C2）。姑息切除后14d每组随机处死6只裸鼠，采用肿瘤转移相关芯片（OHS-028 Oligo GEArray）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。差异基因统计学分析、分组关联度分析及支持向量机（SVM）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，关联向量机（RVM）用于处理回归及分类问题；实时荧光定量聚合酶链反应（FQ—PCR）用于检测mRNA表达。结果由12个基因（BAI1、MTA2、MTA1、SMAD2、GNRH1、CDH8、ITGB3、CHD4、GZMA、ITGA7、CXCR4和TSHR）组成的分子诊断模型对肝癌高、低转移分组的阳性率分别为88．9％和93．5％；MTSS1、转化生长因子-β1（TGF-β1）、SMAD25白细胞介素（IL）-1β及基质金属蛋白酶（MMP）-7基因在高侵袭转移组基因网络中处于核心位置，通过FQ—PCR验证。结论成功构建的分子诊断模型和关键分子可用于评估肝癌转移潜能。
Objective To construct the molecular model and screen the key molecules for evaluation of tumor metastatic potential in nude mice bearing hepatoeellular carcinoma （HCC） xenografts. Methods Orthotopic HCC models were established by implantation of human HCC cell line MHCC97H xeno- grafts with high metastatic potential. Thirty-six nude mice bearing HCC were randomized into three groups 14 days post-operation, including palliative resection group （samples A, B）, sham operation group （sample C1 ） , and control group （ sample C2）. Six mice in each group were sacrificed by cervical dislocation 14 days after palliative resection. Oligo Tumor Metastasis Microarray （ OHS-028 ） and GEArray Expression A- nalysis Suite software were adopted for gene analysis. The methods of support vector machine （ SVM ）, gene significance analysis and gene correlation degree analysis were used to find the markers that could differentiate the metastatic potential of HCC. Gene function net was constructed based on the special gene clustering analysis and multi-dimensional scale. The relevance vector machine （RVM） was used to deal with regression and classification problems. Real-time fluorescence quantitative polymerase chain reaction （FQ-PCR） was applied to detect mRNA expression. Results By the method of support vector machine （SVM）, we found the markers composed of 12 genes （BAIl, MTA2, MTA1, SMAD2, GNRH1, CDH8, ITGB3, CHD4, GZMA, ITGA7, CXCR4, and TSHR） could differentiate the metastatic potential of HCC with positive predictive rate of 88.9% and 93.5% in the groups with high and low metastatic potential respectively. In addition, it was found that MTSS1, transforming growth factor-β1 （TGF-β1）, SMAD2, interleukin-1β （IL-1β） and matrix metalloproteinase-7 （MMP-7） were situated in the central position of the gene function net with high metastatic potential, which was confirmed by Real-time PCR. Conclusion The constructed molecule diagnostic model and the key molecules may be used to evaluate metastatic potential of HCC.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第5期1072-1074,共3页
Chinese Journal of Experimental Surgery
基金
基金项目：国家自然科学基金资助项目（81272401）
上海市卫生局中医药科研基金资助项目（2010L059A）
上海市卫生局中医药科研基金资助项目（2012QJ001A）
关键词
癌
肝细胞
分子模型
侵袭
转移
Carcinoma, hepatocellular
Molecular model
Invasion
Metastasis
