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

1.检测人胰腺癌细胞系(MIA Paca-2、Hs776T、PANC-1、MDA PANC 3、MDA PANC 28、SU86.86、HPAF-II、ASPC-1、Capan-1和HPAC)经裸鼠脾内注射后的肝转移能力。只有四种细胞系(HPAF-II、ASPC-1、CAPAN-1和HPAC)能产生肝转移。在脾内注射后，对这四种细胞系进行体内选择，以增加转移潜能。然而，从转移中生长出来的细胞并没有表现出转移潜能的增加。由于这四种细胞系(HPAF-II、ASPC-1、CAPAN-1和HPAC)在胰腺内移植后被发现转移到肝脏，因此也进行了原位移植的体内选择。高转移性变异体不是通过这种选择产生的。这些结果表明，高转移性胰腺癌…的细胞和分子决定因素的特征更多的细胞应该通过从这四个高转移变异细胞系中选择转移较差的变异来进行。此外，这四个转移细胞与REST(六个细胞系)之间的比较应该被证明是有用的。通过使用表达阵列来比较来自彼此相邻的非侵袭性和高侵袭性肿瘤的一对病理标本的基因表达。有9个基因(MAPkinase5、原癌基因c-src、aggrecan、硫酸软骨素核心蛋白-1、、VisicalIomes、、BM-40、TIMP-1前体、TIMP-1、、)在非侵袭性肿瘤中的表达水平高于癌旁组织。然而，当比较9个病理标本时，在浸润性肿瘤中的表达水平并不总是低的，即非侵袭性和高度恶性的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·费德勒博士的团队建立了人胰腺COLO357癌细胞的高转移变异细胞。其中一种细胞系L3.6pl细胞在裸鼠胰腺原位移植后比亲代FG细胞更易转移。作为一项合作努力，利用Takara智能微阵列检测了这两个细胞之间基因表达水平的差异。33个基因在L3.6pl细胞中的表达水平是亲本FG细胞的3倍以上。较少

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