Liao S K, Perng Y P, Lee L A, Chang K S, Lai G M, Wong E, Ho Y S
Graduate Institute of Clinical Medicine, Chang Gung College of Medicine and Technology, Taoyuan, Taiwan, Republic of China.
Eur J Cancer. 1996 Feb;32A(2):346-56. doi: 10.1016/0959-8049(95)00583-8.
The establishment and characterisation of paired autologous tumour cell line (MST-1) and tumour-infiltrating lymphocyte (TIL) culture from a tumour mass of a 14-year-old Taiwanese girl with soft tissue melanoma are described. MST-1 cells grown in vitro were heterogeneous in morphology, ranging from floating round cells, loosely attached round/oval or elongated cells with prominent pseudopod-like processes, to well-attached spindle and elongated dendritic cells without obvious pseudopods. Immunostaining revealed that major melanoma-associated antigens, such as S100 protein, HMB-45, melanotransferrin, chondroitin sulphate proteoglycan, and the gangliosides GD2 and GD3, were consistently expressed by the tumour tissue, severe combined immunodeficiency (SCID) mouse xenograft and derived cell lines. Flow cytometric analysis of the tumour DNA content showed an index of 1.8 relative to normal peripheral blood lymphocyte DNA. Chromosome analysis revealed all cells at a hypotetraploid level with several clonal chromosome aberrations, including deletions at 10p and 12q, an addition at 12q, translocations t(1;14) and t(5;6). Electron microscopy showed melanosome structures. This observation and the expression of the major melanoma-associated antigens were all indicative of the melanocytic origin of MST-1 tumour. Interleukin-2 (IL-2) expanded TILs had the predominant CD8+ phenotype and the capacity to lyse cells of the cultured autologous tumour. The availability of the soft tissue melanoma cell line, the SCID mouse xenograft tumour system as well as autologous TILs described herein would provide useful materials for identifying T-cell-defined antigens as well as a model system for devising individualised cancer biotherapeutic strategies. This cell line can also be used for further studies aimed at uncovering the histogenesis of this rare cancer.
本文描述了从一名患有软组织黑色素瘤的14岁台湾女孩的肿瘤块中建立配对的自体肿瘤细胞系（MST-1）和肿瘤浸润淋巴细胞（TIL）培养物并对其进行表征的过程。体外培养的MST-1细胞形态各异，包括漂浮的圆形细胞、松散附着的圆形/椭圆形或带有明显伪足样突起的细长细胞，以及牢固附着的纺锤形和细长树突状细胞，无明显伪足。免疫染色显示，主要的黑色素瘤相关抗原，如S100蛋白、HMB-45、黑素转铁蛋白、硫酸软骨素蛋白聚糖以及神经节苷脂GD2和GD3，在肿瘤组织、严重联合免疫缺陷（SCID）小鼠异种移植物和衍生细胞系中均持续表达。肿瘤DNA含量的流式细胞术分析显示，相对于正常外周血淋巴细胞DNA，其指数为1.8。染色体分析显示所有细胞处于亚四倍体水平，有几个克隆性染色体畸变，包括10p和12q缺失、12q增加、易位t(1;14)和t(5;6)。电子显微镜显示有黑素小体结构。这一观察结果以及主要黑色素瘤相关抗原的表达均表明MST-1肿瘤起源于黑素细胞。白细胞介素-2（IL-2）扩增的TILs具有主要的CD8+表型，并具有裂解培养的自体肿瘤细胞的能力。本文所述的软组织黑色素瘤细胞系、SCID小鼠异种移植物肿瘤系统以及自体TILs的可用性，将为鉴定T细胞定义的抗原提供有用材料，并为设计个体化癌症生物治疗策略提供模型系统。该细胞系还可用于进一步研究，以揭示这种罕见癌症的组织发生机制。
