Establishment of hepatic metastasis model of pancreatic carcinoma and it application to the development of therapeutic agents

1. 在胸腺裸鼠脾内注射人胰腺癌细胞系（MIA PaCa-2、Hs776T、Panc -1、MDA Panc 3、MDA Panc 28、SU86.86、HPAF-II、AsPC-1、Capan-1和HPAC）后，检测其肝转移能力。只有四种细胞系（hpf - ii, AsPC-1， Capan-1和HPAC）显示产生肝转移。这四种细胞系在脾内注射后进行了体内选择，以增加转移潜力。然而，从转移瘤中生长出来的细胞没有显示出转移潜力的增加。由于这四种细胞系（haff - ii、AsPC-1、Capan-1和HPAC）在胰腺内移植后会转移到肝脏，因此我们也进行了原位移植的体内选择。高度转移的变异，不是通过这种选择产生的。这些结果表明了高转移性胰腺癌的细胞和分子决定因素的特征，应该通过从这四种高转移性变异细胞系中选择低转移性变异来进行更多的细胞研究。此外，我们建议将这四种转移细胞与其他（六种细胞系）进行比较应该证明是有用的。采用表达阵列技术，比较非侵袭性和高度侵袭性相邻肿瘤病理标本的基因表达情况。9个基因（MAPkinase kinase-5、原癌基因c-src、aggrecan 1、硫酸软骨素核心蛋白-1、Visican同分异构体、BM-40、血栓响应蛋白1前体、TIMP-1、前胶原c蛋白）在非浸润性肿瘤中的表达水平高于邻近浸润性肿瘤。然而，在9个病理标本中，浸润性肿瘤的表达水平并不总是低的，即非浸润性和高度恶性胰腺癌细胞之间的差异表达并不总是如此。德克萨斯大学安德森癌症中心的以赛亚·j·菲德勒博士小组建立了人类胰腺COLO357癌细胞的高度转移变异细胞。其中一种细胞系L3.6pl细胞在原位移植到裸鼠胰腺后比亲本FG细胞更具转移性。通过合作，我们用Takara智能芯片检测了这两种细胞之间基因表达水平的差异。有33个基因在L3.6pl细胞中的表达量比亲本FG细胞高出3倍以上。少

1. Human pancreatic carcinoma cell lines (MIA PaCa-2, Hs776T, PANC-1, MDA Panc 3, MDA Panc 28, SU86.86, HPAF-II, AsPC-1, Capan-1, and HPAC) were tested for their capacity to generate hepatic metastataic following intrasplenic injections in athymic nude mice. Only four cell line (HPAF-II, AsPC-1, Capan-1, and HPAC) were shown to produce hepatic metastasis. These four cell lines were subjected to in vivo selections for increased metastatic potentials following intrasplenic injections. However, cells grown out from metastases did not show increase in the metastatic potential.2. Because these four cell lines (HPAF-II, AsPC-1, Capan-1, and HPAC) were shown to metastasize to the liver after intra-pancreatic transplantations, in vivo selections with the orthotopic transplantation were also conducted. Highly metastatic variants, were not generated by this selection.3. These results indicated that characterization of cellular and molecular determinants of highly metastatic pancreatic carcinoma … More cells should be conducted through selections of poorly metastatic variants from these four highly metastatic variant cell lines. Also, it is suggested that comparisons between these four metastaic cells and rest (six cell lines) should prove to be useful.4. A pair of pathological specimens from noninvasive and highly invasive tumors adjacent to each other was compared for the gene expression by the use of expression arrays. Nine genes (MAPkinase kinase-5、 protooncogene c-src、 aggrecan 1、chondroitin sulfate coreprotein-1、Visican isohomes、BM-40、thrombospondin-1 precursor、TIMP-1、procollagen C-protein) were identified to be higher in the expression level in the non-invasive tumor than in the adjacent invasive tumor. However, the levels were not always low in the invasive tumors when nine pathological specimens were compared, i.e. the differential expression between non-invasive and highly malignant pancreatic carcinoma cells was not always the case.5. Dr. Isaiah J. Fidler's group at the University of Texas M. D. Anderson Cancer Center established highly metastatic variant cells of human pancreatic COLO357 carcinoma cells. One of these cell lines, L3.6pl cells, was shown to be more metastatic after orthotopic transplantation into pancreas of nude mice than the parental FG cells. As a collaborative effort, difference in the level of gene expression between these two cells was examined by Takara Intelligene Microarray. Thirty three genes were found to be expressed more than three times higher in L3.6pl cells as compared to parental FG cells. Less