Assay of anti-cancer drugs in tissue culture: conditions affecting their ability to incorporate 3H-leucine after drug treatment.
Assay of anti-cancer drugs in tissue culture: conditions affecting their ability to incorporate 3H-leucine after drug treatment.
复制标题
组织培养中抗癌药物的测定：药物治疗后影响其吸收 3H-亮氨酸能力的条件。
作者：
R. Freshney;J. Paul;I. Al;Kane
我们已经尝试构建一种可能适用于人类肿瘤原代培养物的测定方法，以测量暴露于抗肿瘤药物后呈指数增长的单层培养物的存活率。用去除药物后细胞的蛋白质合成能力来评估细胞存活。采用HeLa细胞来避免人类肿瘤原代培养物的成分变异性和异质性，并使用微滴定托盘构建了一种试验，以提供大量的重复培养物，而不需要大量的细胞。在几乎所有被检查的病例中，暴露于药物的持续时间的增加增加了敏感性。同样，去除药物后培养时间的增加会增加对烷基化剂的敏感性，但可以从暴露于某些周期依赖性药物中恢复。一些使用的药物在培养条件下被证明是不稳定的，长春花碱被积极代谢，尽管这种不稳定性不一定反映在药物作用的时间过程中。当细胞密度增加到一定程度时，3h -胸腺嘧啶掺入的密度限制变得明显，Mustine的敏感性就会降低。文中还讨论了这些变化以及减少其影响的可能方法。
An attempt has been made to construct an assay potentially suitable for use with primary cultures of human tumours to measure the survival of exponentially growing monolayer cultures after exposure to anti-neoplastic drugs. Cell survival was assessed using their protein synthetic capacity after removal of drugs. HeLa cells were employed to avoid the ingerent variability and heterogeneity of primary cultures from human tumours, and an assay has been constructed using microtitration trays to provide large numbers of replicate cultures without the requirement of a large number cells. An increase in the duration of the exposure to drug increased sensitivity in nearly all cases examined. Similarly, an increase in the period of culture following drug removal produced increased sensitivity to alkylating agents but allowed recovery from exposure to certain cycle-dependent drugs. Some of the drugs used were shown to be unstable under culture conditions and vinblastine was actively metabolized, although this instability was not necessarily reflected in the time course of the drug's effect. Mustine sensitivity was shown to be reduced by an increase in cell density at a level where density limitation of 3H-thymidine incorporation becomes apparent. These variations and possible methods of minimizing their effects are discussed.