摘要
目的:筛选肝细胞癌(hepatocellular carcinoma,HCC)中表达水平较正常肝细胞差异大、特异性好的基因,建立肝癌分子诊断指数,以期从分子病因学角度实现对肝癌的早期诊断.方法:随机选择38例手术切除组织标本,其中正常肝5例、肝炎4例、肝硬化(LC)12例、HCC和癌旁组织17例,实时荧光定量RT-PCR检测9个基因的表达,以管家基因G3PDH为对照,2-△△CT法计算目的基因相对表达量,依据表达异常的基因个数建立分子诊断指数.结果:GPC3等6个基因在正常肝、肝炎、LC与HCC组的两组或多组间存在差异(P<0.05);GPC3、E2F1从肝炎组到LC、HCC组呈递增趋势,HGF、CLDN10在LC组表达量升高,而在HCC组明显下降,PTEN、PRDM2、MGMT从肝炎组到LC、HCC组呈递减趋势;9个基因在LC组分子诊断指数平均值为1.58,在HCC组平均值为5.24,二者差异显著;GPC3、E2F1、MMP2从癌旁到癌呈递增趋势,CLDN10、HGF、PTEN、DLC1、PRDM2、MGMT从癌旁到癌呈递减趋势.结论:通过筛选更多基因的组合分析,分子诊断指数有望成为鉴别诊断早期肝癌的一种有效方法.
AIM： To develop molecular diagnostic indexes to detect hepatocellular carcinoma （HCC） using genes whose expression level differs markedly from the normal tissue. METHODS： Thirty eight tissue samples were obtained from surgical resection at random in- cluding 5 normal liver, 4 hepatitis, 12 liver cirrhosis, 17 HCC and paracancer tissues. Gene expression in 9 cases was detected using real-time fluorescent qualification RT-PCR on the control of G3PDH （housekeeping gene）. The method of 2^-△△CT was used to calculate the relative qualification of target genes, and then the molecular diagnostic index was estimated according to the gene numbers of abnormal expression. RESULTS： Six gene expression differed among normal, hepatitis, LC and HCC groups. GPC3 and E2F1 showed an increasing trend, while PTEN, PRDM2, and MGMT showed a decreasing trend. But HGF, CLDN10 expression was elevated in LC, decreased in HCC. For 9 genes as molecular diagnosis index, their average value was 1.58 in LC, 5.24 in HCC, and the difference was significant. GPC3, E2F1 and MMP2 increased gradually from paracancer to cancer. CLDN10, HGF, PTEN, DLC1, PRDM2, and MGMT decreased gradually from paracancer to cancer. CONCLUSION： The method of molecular diagnostic index can be potentially used to detect and differentiate early HCC effectively by analyzing the combination of more screening genes.
出处
《世界华人消化杂志》
CAS
北大核心
2009年第10期985-991,共7页
World Chinese Journal of Digestology
基金
天津市科委攻关基金资助项目
No.05YFSZSF02500~~
关键词
肝细胞癌
基因
表达
实时荧光定量RT—PCR
分子诊断指数
Hepatocellular carcinoma
G-ene expression
Expression
Real-time fluorescent quantitation polymerase chain reaction
Molecular diagnostic index
