CAJ | 학술논문

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,14 d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14 d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)、SMAD2(*)、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.
목적 구건간세포암(간암)전이상관분자모형병사선관건분자,용우평고간암전이잠능.방법 이고전이인간암세포MHCC97H건립라서원위이식류모형,14 d후장36지성모라서수궤분성고식성간절제조(획취간암표본A、B)、가수술조(획취간암표본C1)화대조조(획취간암표본C2).고식절제후14 d매조수궤처사6지라서,채용종류전이상관심편(OHS-028 Oligo GEArray)검측간암기인표체,사용종합형GEArray분석배투연건분석심편수거.차이기인통계학분석、분조관련도분석급지지향량궤(SVM)용우사선종류전이상관표지기인；응용기우특정성치병기인적기인취류화다유량표구건기인공능망락,관련향량궤(RVM)용우처리회귀급분류문제；실시형광정량취합매련반응(FQ-PCR)용우검측mRNA표체.결과 유12개기인(BAI1、MTA2、MTA1、SMAD2、GNRH1、CDH8、ITGB3、CHD4、GZMA、ITGA7、CXCR4화TSHR)조성적분자진단모형대간암고、저전이분조적양성솔분별위88.9％화93.5％；MTSS1、전화생장인자-β1(TGF-β1)、SMAD2(*)、백세포개소(IL)-1β급기질금속단백매(MMP)-7기인재고침습전이조기인망락중처우핵심위치,통과FQ-PCR험증.결론 성공구건적분자진단모형화관건분자가용우평고간암전이잠능.
Objective To construct the molecular model and screen the key molecules for evaluation of tumor metastatic potential in nude mice bearing hepatocellular carcinoma (HCC) xenografts.Methods Orthotopic HCC models were established by implantation of human HCC cell line MHCC97H xenografts 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 Analysis 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 (BAI1,MTA2,MTA1,SMAD2,GNRH1,CDHS,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-1 β (IL-1 β) 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.