Fujii Y, Itoyanagi H, Saegusa Y, Kuro-o M, Matsuda Y, Shiroko Y, Eriguchi M, Hasumi K
Department of Surgery, Institute of Medical Science Hospital, University of Tokyo.
Jpn J Cancer Res. 1993 Oct;84(10):1055-61. doi: 10.1111/j.1349-7006.1993.tb02801.x.
The objective of this study was to examine the identity and characteristics of a spontaneously occurring murine retroperitoneal tumor of BALB/c mouse origin that selectively metastasized to the liver. From the primary tumor, a permanent cell line, termed LMFS (liver metastasis from sarcoma) was established in vivo and in vitro. After a subcutaneous injection of more than 1 x 10(5) cells in the side back of mice, the LMFS cells proliferated at the inoculation site (100% take) and induced metastatic nodules spontaneously in the liver, but not in the lung. By the limiting dilution technique, a cloned cell line, LMFS-1, was established in vitro. The LMFS-1 cell line had similar morphological characteristics to the LMFS cells both in vitro and in vivo. The doubling time of the LMFS-1 cell line was 10 h in passage 60. The number of chromosomes ranged from 71 to 108 and 93% of metaphases showed near-tetraploidy. In microscopic examination, no specific arrangement of the LMFS tumor cells was seen; the LMFS cell had medium- to large-sized atypical nuclei and clear and large cytoplasm. Electronmicroscopy showed that the cytoplasm of the LMFS cell had a moderate amount of rough-surfaced endoplasmic reticulum but no desmosomes or microvilli. Immunohistochemically, the LMFS cells were positive for vimentin, but showed no reaction for keratin or cytokeratin. Therefore, the LMFS tumor was considered to be an undifferentiated sarcoma. The LMFS cell line should be a useful tool not only for studies of metastasis, but also for experiments on the therapy of hepatic tumors.
本研究的目的是检测一种自发产生的、起源于BALB/c小鼠的腹膜后肿瘤的特征及特性,该肿瘤可选择性转移至肝脏。从原发性肿瘤中,在体内和体外建立了一个永久性细胞系,称为LMFS(肉瘤肝转移细胞系)。在小鼠背部皮下注射超过1×10⁵个细胞后,LMFS细胞在接种部位增殖(接种成功率100%),并自发在肝脏而非肺中诱导形成转移结节。通过有限稀释技术,在体外建立了一个克隆细胞系LMFS-1。LMFS-1细胞系在体外和体内均具有与LMFS细胞相似的形态特征。LMFS-1细胞系在第60代时的倍增时间为10小时。染色体数目在71至108条之间,93%的中期细胞显示近四倍体。在显微镜检查中,未观察到LMFS肿瘤细胞的特定排列;LMFS细胞具有中等至大型的非典型细胞核以及清晰且较大的细胞质。电子显微镜显示,LMFS细胞的细胞质含有适量的粗面内质网,但没有桥粒或微绒毛。免疫组织化学检测显示,LMFS细胞波形蛋白呈阳性,但角蛋白或细胞角蛋白无反应。因此,LMFS肿瘤被认为是一种未分化肉瘤。LMFS细胞系不仅应是转移研究的有用工具,也是肝肿瘤治疗实验的有用工具。