Investigation by micro array of genes associated with hepatocellular carcinoma

我们利用DNA微阵列检测了4例肝部分切除后早期异位复发和无异位复发的肝癌患者的差异表达基因。在获得知情同意后，对四例肝细胞癌切除的肝组织进行基因表达分析。在一周内，使用Atlas^&lt；TM&gt；Glass总RNA分离试剂盒(Clontech，#K1036-1)从100-200毫克的肝组织中提取总RNA。从20μg的核糖核酸中提取c DNA，用Atlas Glass荧光标记试剂盒进行荧光染料标记。将标记探针的80个μL与Atlas^&lt；TM&gt；Glass Array1.0杂交。基因的表达具有一定的比率(癌症/非癌症)。3/1被认为是上调，而1/3被归类为下调。在4种肿瘤中表达显著上调的基因分别是MAP/微管亲和力调节激酶3、HGF受体KiSS-1、MMP10、p34蛋白激酶、CDK4、Ien-Gamma受体β亚基、血管生成素1受体、内皮细胞激酶转录延伸因子SII、PAX3、神经钙粘素、CXC趋化因子、Ien-Gamma、NCAM-1和MMP2。下调的基因有：基质金属蛋白酶-8、腺苷环化酶VII、信号素E、转化生长因子-β受体1、Mark-3、LCAT、干扰素调节因子1、NK-1受体、转录因子reIIB、白介素15和肿瘤坏死因子α转换酶。基质金属蛋白酶-2、基质金属蛋白酶-8和肿瘤坏死因子-α转换酶目前被认为是根据肿瘤复发而差异表达的候选基因。我们得出的结论如下：这些肿瘤的异位复发可能与某些基因有关。

We utilized a DNA micro array and examined genes expressed differentially in 4 cases with and without early ectopic recurrence of HCC following partial hepatectomy. After informed consent was obtained, resected liver tissue from four HCC cases was analyzed for gene expression. Within a week, total RNA was extracted from 100 to 200 mg liver tissue using the Atlas^<TM> Glass Total RNA Isolation Kit (Clontech, #K1036-1). After cDNA was prepared from 20 μg of RNA, it was labeled with fluorescent dye using the Atlas Glass Fluorescent Labeling Kit. Eighty μl of the labeled probe was hybridized to the Atlas^<TM> Glass Array 1.0. Genes expressed with a ratio (cancer / non-cancer). 3/1 were considered, to be up-regulated, while those = 1/3 were classified as down-regulated. The markedly up-regulated genes in the four tumors were Map/microtubule affinity-regulating kinase 3, HGF receptor KiSS-1, MMP-10, p34 protein kinase, CDK4, IEN gamma receptor beta subunit, angiopoietin 1 receptor, endothelial cell kinase transcription elongation factor SII,PAX3, neural cadherin, CXC chemokain, IEN gamma, NCAM-1, and MMP-2. The down-regulated genes were MMP-8, adenylate cyclase VII, semaphorin E, TGF-beta receptor type 1, MARK-3,LCAT,IFN regulatory factor 1, Nk-1 receptor, transcription factor reIIB, interleukin 15, and TNF alpha converting enzyme. MMP-2, MMP-8, and TNF-alpha converting enzyme are currently being examined as candidate genes differentially expressed depending on the recurrence of the tumor. We concluded as follows, the possibility is suggested that there are some genes involved in the ectopic recurrence of these tumors.