Identification of a novel tumor marker(s), based on the comprehensive analysis of genes expressed selectively in micometastais site.

1)具有丝氨酸/苏氨酸激酶活性的原癌基因的异常表达；皮姆-3，内胚层来源器官的恶性病变。采用荧光差异显示法比较小鼠肝癌发生模型中癌前病变和恶性病变的基因表达模式。结果，我们观察到具有丝氨酸/苏氨酸激酶活性的原癌基因Pim-3在癌前病变和恶性病变中异常表达，而在正常肝组织中不表达。在人肝细胞癌（HCC）和正常肝组织中也得到了类似的观察结果。Pim-3在人肝癌细胞系中组成性表达，短干扰RNA抑制Pim-3的表达可诱导人肝癌细胞系凋亡。Pim-3在胰腺、结肠等其他内胚层源性器官中未检出，但在这些器官的癌前病变和恶性病变中表达增强。在人胰腺癌和结直肠癌细胞系中，Pim-3和一个促凋亡分子Bad均组成性表达，Bad与Pim共定位，并在112-Ser位点磷酸化，这代表了Pim的失活形式。短干扰RNA抑制Pim-3表达，可降低Bad在112-Ser位点的磷酸化水平，降低抗凋亡分子Bcl-XL的表达，最终促进细胞凋亡。这些观察结果表明，Pim-3的异常表达可以通过抑制细胞凋亡促进内胚层来源器官（包括肝脏、胰腺和结肠）的癌变，并且Pim-3可以作为一种新的肿瘤标志物来检测这些器官的癌前病变和恶性病变。我们观察到趋化因子CCL3及其受体CCR1在二乙基亚硝胺诱导的小鼠肝癌发生的早期阶段可以抑制癌变，而CCL3-CCR1轴可以通过诱导血管生成促进相同肝癌发生的癌变。我们进一步证明了另一种趋化因子CCR2及其受体CCR2可以通过激活肝星状细胞（Ito细胞）并最终加速新血管形成来促进肝转移。少

1)Aberrant expression of a proto-oncogene with serine/threonine kinase activity; Pim-3, in malignant lesions of endoderm-derived organs. We compared gene expression patterns in pre-malignant and malignant lesions in murine hepatocarinogenesis model, by using fluorescent differential display method. As a result, we observed that a proto-oncogene with serine/threonine kinase activity, Pim-3 was aberrantaly expressed in pre-malignant and malignant lesions, but not normal hepatic tissues. Similar observations were obtained also on human hepatocellular carcinoma (HCC) and normal liver tissues. Pim-3 was constitutively expressed in human HCC cell lines and the inhibition of Pim-3 expression by short interfering RNA induced the apoptosis of human HCC cell lines. Pim-3 was not detected in other endoderm-derived organs such as pancreas and colon, but its expression was enhanced in the pre-malignant lesions and malignant lesions of these organs. In both human pancreatic cancer and colorectal can … More cer cell lines, Pim-3 was constitutively expressed and a pro-apoptotic molecule, Bad was co-localized with Pim and was phosphorylated at 112-Ser, which represents its inactive form. The inhibition of Pim-3 expression by short interfering RNA reduced the phosphorylation of Bad at 112-Ser and the expression of an anti-apoptotic molecule, Bcl-XL and eventually enhanced the apoptosis. These observations suggest that aberrant expression of Pim-3 can contribute to carcinogenesis in endoderm-derived organs, including liver, pancreas, and colon, by inhibiting apoptosis and that Pim-3 can be a novel tumor marker to detect pre-malignant and malignant lesions of these organs.2)Roles of chemokines and hepatocarcinogenesis and liver metastasis We observed that a chemokine, CCL3 and its receptor, CCR1, can inhibit the carcinogenesis in the early phase of murine hepatocarcinogenesis induced by diethylnitrosamine administration but that the CCL3-CCR1 axis can promote carcinogenesis of the same hepatocarcinogenesis, by inducing angiogenesis. We further demonstrated that another chemokine, CCL2 and its receptor, CCR2, can promote liver metastasis by activating hepatic stellate cells (Ito cells) and eventually accelerating neovascularization. Less