Klostergaard J
Lymphokine Res. 1985 Fall;4(4):309-17.
Pretreatment of the murine mammary adenocarcinoma cell line, EMT-6, with low levels (0.8-1.2 micrograms/ml) of actinomycin D, prior to incubation with a spectrum of cytotoxic cytokines, converted this target from marked resistance to extreme sensitivity. The drug-treated EMT-6 was from 5-50 fold more sensitive to cytokine attack than the widely used actinomycin D-treated murine L-929 target. Drug-induced growth inhibition allowed evaluation solely of the cytolytic effects of these cytokines. Lysis was evaluated after a 16-24 hr incubation by the uptake by viable cells of neutral red or by their reduction of [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. This microcytotoxicity assay should facilitate purification and characterization of cytotoxic cytokines, the purification of their mRNAs in translation systems and the detection of the expression of their encoding genes.
在用一系列细胞毒性细胞因子孵育之前，先用低水平（0.8 - 1.2微克/毫升）的放线菌素D预处理小鼠乳腺腺癌细胞系EMT - 6，可使该靶细胞从显著抗性转变为极度敏感。经药物处理的EMT - 6对细胞因子攻击的敏感性比广泛使用的经放线菌素D处理的小鼠L - 929靶细胞高5至50倍。药物诱导的生长抑制使得能够仅评估这些细胞因子的细胞溶解作用。在16 - 24小时孵育后，通过活细胞对中性红的摄取或其对[3 - （4,5 - 二甲基噻唑 - 2 - 基） - 2,5 - 二苯基溴化四氮唑]的还原作用来评估细胞溶解。这种微量细胞毒性测定法应有助于细胞毒性细胞因子的纯化和表征、其mRNA在翻译系统中的纯化以及其编码基因表达的检测。
