Ogawa K, Nakanishi H, Takeshita F, Futakuchi M, Asamoto M, Imaida K, Tatematsu M, Shirai T
First Department of Pathology, Nagoya City University, Medical School, 1 Kawasumi, Mizuho-cho, Nagoya 467-8601, Japan.
Int J Cancer. 2001 Mar 15;91(6):797-802. doi: 10.1002/1097-0215(200002)9999:9999<::aid-ijc1140>3.0.co;2-#.
For better understanding of cancer metastasis, we have established an in vivo model for induction of highly metastatic hepatocellular carcinomas (HCC) in male F344 rats. From 1 tumor, 4 cell lines with differing metastatic potential (C1, C2, C6, C5F) were established by subcloning using the limited-dilution cloning technique. Two other lines, N1 and L2, arose from another primary HCC and a lung metastatic lesion, respectively. Although cell adhesion of each cell line in culture medium was different, tumors developing in the subcutis of nude mice after transplantation were all moderately differentiated HCC with a trabecular pattern. On subcutaneous injection into nude mice, all 6 cell lines proved to be tumorigenic in the injection site and C5F was highly metastatic to the lung. With injection into the tail vein, N1 and L2 formed frequent metastases in the lung as well as in lymph nodes. Using intraperitoneal injection, C1, C6, N1 and L2 showed marked disseminated growth in the abdominal cavity with bloody ascitis. Northern blot analysis revealed expression of known metastasis-related genes, KAI1 and heparanase, to be decreased in C5F, but no differences in expression of nm23-H1 were evident. A point mutation in the GSK-3beta phosphorylation site of the beta-catenin gene was found in L2. These transplantable HCC cell lines that have different metastatic ability should be useful for elucidation of mechanisms of metastasis.
为了更好地理解癌症转移，我们建立了一种在雄性F344大鼠中诱导高转移性肝细胞癌（HCC）的体内模型。从1个肿瘤中，使用有限稀释克隆技术通过亚克隆建立了4种具有不同转移潜能的细胞系（C1、C2、C6、C5F）。另外两个细胞系N1和L2分别来自另一个原发性HCC和一个肺转移病灶。尽管每个细胞系在培养基中的细胞黏附情况不同，但移植后在裸鼠皮下形成的肿瘤均为中等分化的小梁状HCC。皮下注射到裸鼠体内后，所有6种细胞系在注射部位均具有致瘤性，且C5F对肺具有高转移性。尾静脉注射后，N1和L2在肺和淋巴结中频繁形成转移灶。腹腔注射后，C1、C6、N1和L2在腹腔内表现出明显的播散性生长并伴有血性腹水。Northern印迹分析显示，已知的转移相关基因KAI1和乙酰肝素酶在C5F中的表达降低，但nm23-H1的表达没有明显差异。在L2中发现β-连环蛋白基因的GSK-3β磷酸化位点存在点突变。这些具有不同转移能力的可移植HCC细胞系应有助于阐明转移机制。
