Melber K, Zhu G, Diamond L
Wistar Institute, Philadelphia, Pennsylvania 19104.
Cancer Res. 1989 Jul 1;49(13):3650-5.
Normal human melanocytes, which require the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) for growth in culture, were transfected with a SV40 T-antigen-containing plasmid by using the technique of electroporation, and were scored for colonies of morphologically altered cells. The frequency of transformed colonies was higher when selection was done in the absence rather than the presence of TPA in the medium. Three cell lines derived from transformed colonies were characterized. All show an enhanced growth rate compared to parental cells, anchorage independence, loss of dependence on medium supplements for growth, chromosomal abnormalities, an extended life span, and growth inhibition by TPA. They express nerve growth factor receptor and Mr 97,000 protein, melanotransferrin, two antigens usually associated with melanocytic cells, but the transformed cells are not pigmented. The three cell lines underwent crisis at about passage 10 posttransfection; one cell line recovered and appears to have unlimited growth potential. None of the cell lines is tumorigenic. They should be interesting models for studying multistage carcinogenesis in human cells and transcriptional activation by TPA.
正常人类黑素细胞在培养中生长需要佛波酯12 - O - 十四烷酰佛波醇-13 - 乙酸酯（TPA），通过电穿孔技术用含SV40 T抗原的质粒转染这些细胞，并对形态改变的细胞集落进行计数。当在培养基中不存在而非存在TPA的情况下进行选择时，转化集落的频率更高。对源自转化集落的三个细胞系进行了表征。与亲代细胞相比，所有细胞系均显示出生长速率提高、不依赖贴壁生长、生长不依赖培养基补充物、染色体异常、寿命延长以及对TPA生长抑制敏感。它们表达神经生长因子受体和97000 Mr蛋白、黑素转铁蛋白，这两种抗原通常与黑素细胞相关，但转化细胞无色素沉着。这三个细胞系在转染后约第10代时经历危机；一个细胞系恢复并似乎具有无限生长潜能。这些细胞系均无致瘤性。它们应是研究人类细胞多阶段致癌作用和TPA转录激活的有趣模型。
