Ding J L, Tan B C
Department of Zoology, National University of Singapore, Kent Ridge.
Cytobios. 1989;59(237):101-14.
Two methods of DNA-mediated gene-transfer, namely, CaPO4 co-precipitation and polybrene/DMSO-induction were used to analyse oncogenes from various cancer cell-lines. Third time transfection of mouse fibroblast cells, NIH/3T3 with 10, 100, 1,000 and 3,000 ng of DNA from spontaneous human hepatoma cells produced 41, 4, 45 and 56 foci, respectively. It appears that 100 ng of this hepatoma cell DNA gave the least effective transformation ability. However, with two spontaneous mammary cancer cell lines, 100 ng DNA gave the highest number of foci. In general, during the initial transfection, the cells appeared semi-transformed, showing stellate growth patterns. With repeated transfections they gave way to foci. However, with aflatoxin-induced rat hepatoma cell (JB-1) DNA, only stellate growths persisted to indicate intermediate transformation. Transfection of the epithelial-like normal rat liver cell line, BL8 with JB-1 DNA produced haphazard growth pattern, although a hepatoma-associated enzyme, gamma-glutamyl transferase was introduced into the BL8 cells which previously had no detectable enzyme activity.
采用两种DNA介导的基因转移方法，即磷酸钙共沉淀法和聚凝胺/二甲基亚砜诱导法，对来自各种癌细胞系的癌基因进行分析。用来自人自发性肝癌细胞的10、100、1000和3000 ng DNA对小鼠成纤维细胞NIH/3T3进行第三次转染，分别产生了41、4、45和56个集落。看来100 ng这种肝癌细胞DNA的转化能力最低。然而，对于两个自发性乳腺癌细胞系，100 ng DNA产生的集落数量最多。一般来说，在初次转染期间，细胞呈现半转化状态，显示出星状生长模式。随着转染次数的增加，它们被集落取代。然而，用黄曲霉毒素诱导的大鼠肝癌细胞（JB-1）DNA转染时，只有星状生长持续存在，表明处于中间转化状态。用JB-1 DNA转染上皮样正常大鼠肝细胞系BL8，产生了杂乱的生长模式，尽管一种与肝癌相关的酶——γ-谷氨酰转移酶被导入了之前没有可检测酶活性的BL8细胞中。
