Investigation by micro array of genes associated with hepatocellular carcinoma

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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.