Heo D S, Snyderman C, Gollin S M, Pan S, Walker E, Deka R, Barnes E L, Johnson J T, Herberman R B, Whiteside T L
Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania.
Cancer Res. 1989 Sep 15;49(18):5167-75.
Twenty-one head and neck squamous cell carcinoma (HNSCC) cell lines were established from 89 fresh tumor specimens in order to study the biology of HNSCC lines, establish tumors in nude mice, and evaluate the sensitivity to immunological effector cells of these tumors in vitro and in vivo in nude mice. The lines were established from explants using differential trypsinization and culture for 2 to 20 mo. The explants were derived from 11 different sites. Three pairs of lines were derived from both the primary tumor and metastatic lymph nodes in the same patients. All cultures grew as either compact or diffuse adherent monolayers, and they had a median doubling time of 86 h (range, 33 to 531 h). DNA fingerprinting confirmed that the HNSCC lines were individual isolates. Thirteen of 14 lines tested induced tumors in athymic mice. The histology of each line growing in nude mice was similar to that of the original tumor tissue. Immunocytochemistry showed keratin production in all lines tested. Aneuploidy (36 to 87 chromosomes) was present in all 16 lines studied; the median chromosome number for lines derived from primary tumors was 70, whereas for lines originating from metastatic or recurrent tumors, it was 54. Karyotypic analysis showed deletion of the short arm of chromosome 3 (3p-) in 12 of 16 cell lines and trisomy 6 in 12 of 16 lines. In addition, translocations between chromosomes 9 and 11 or 9 and 12 were each present in five of 16 lines tested. The HNSCC lines were resistant to lysis by natural killer cells, but were efficiently lysed by lymphokine-activated killer cells in 4-h 51Cr release assays. These new lines have allowed us to establish a model of local adoptive immunotherapy of HNSCC in tumor-bearing nude mice, and they provide a resource for future studies of the biology of HNSCC.
为了研究头颈部鳞状细胞癌(HNSCC)细胞系的生物学特性、在裸鼠体内建立肿瘤模型以及评估这些肿瘤在体外和裸鼠体内对免疫效应细胞的敏感性,从89份新鲜肿瘤标本中建立了21种HNSCC细胞系。这些细胞系通过差异胰蛋白酶消化法从外植体建立,并培养2至20个月。外植体来自11个不同部位。三对细胞系来自同一患者的原发性肿瘤和转移性淋巴结。所有培养物均以紧密或弥漫性贴壁单层生长,其平均倍增时间为86小时(范围为33至531小时)。DNA指纹图谱证实这些HNSCC细胞系是独立的分离株。14个测试细胞系中有13个能在无胸腺小鼠中诱导肿瘤。在裸鼠体内生长的每个细胞系的组织学与原始肿瘤组织相似。免疫细胞化学显示所有测试细胞系均产生角蛋白。所研究的16个细胞系均存在非整倍体(36至87条染色体);源自原发性肿瘤的细胞系的中位数染色体数为70,而源自转移性或复发性肿瘤的细胞系的中位数染色体数为54。核型分析显示,16个细胞系中有12个存在3号染色体短臂缺失(3p-),16个细胞系中有12个存在6号染色体三体。此外,在16个测试细胞系中有5个分别存在9号和11号或9号和12号染色体之间的易位。这些HNSCC细胞系对自然杀伤细胞的裂解具有抗性,但在4小时51Cr释放试验中能被淋巴因子激活的杀伤细胞有效裂解。这些新的细胞系使我们能够在荷瘤裸鼠中建立HNSCC局部过继免疫治疗模型,并为未来HNSCC生物学研究提供了资源。