Investigation by micro array of genes associated with hepatocellular carcinoma

我们利用DNA微阵列检测了4例肝部分切除术后早期异位复发和未异位复发的HCC患者的基因表达差异。在获得知情同意后，分析了来自四个HCC病例的切除的肝组织的基因表达。在一周内，使用Atlas ® <TM>Glass总RNA分离试剂盒（Clontech，#K1036-1）从100至200 mg肝组织中提取总RNA。从20 μg RNA制备cDNA后，使用Atlas Glass Fluorescent Labeling Kit用荧光染料标记。将80 μl标记探针与Atlas ® <TM>Glass Array 1.0杂交。以比率（癌症/非癌症）表达的基因。3/1被认为是上调的，而那些= 1/3被归类为下调的。在4种肿瘤中显著上调的基因是Map/微管亲和力调节激酶3、HGF受体KiSS-1、MMP-10、p34蛋白激酶、CDK 4、IEN γ受体β亚单位、血管生成素1受体、内皮细胞激酶转录延伸因子SII、PAX 3、神经钙粘蛋白、CXC趋化因子、IEN γ、NCAM-1和MMP-2。下调的基因是MMP-8、腺苷酸环化酶VII、脑信号蛋白E、TGF-β 1型受体、MARK-3、LCAT、IFN调节因子1、Nk-1受体、转录因子reIIB、白细胞介素15和TNF α转化酶。MMP-2、MMP-8和TNF-α转化酶目前正在作为根据肿瘤复发而差异表达的候选基因进行研究。我们的结论如下，可能有一些基因参与了这些肿瘤的异位复发。

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.