Intracellular uptake of 7-con-o-methylnogarol and adriamycin by cells in culture and its relationship to cell survival.
Intracellular uptake of 7-con-o-methylnogarol and adriamycin by cells in culture and its relationship to cell survival.
复制标题
培养细胞对 7-con-o-methylnogarol 和阿霉素的细胞内摄取及其与细胞存活的关系。
作者：
B. Bhuyan;J. Mcgovren;S. Crampton
摘要7-酮-O-甲基橙花酚(7-omen)是一种新的抗肿瘤药物，对小鼠P388、L1210白血病(寿命延长140%)和B16黑色素瘤(寿命延长114%)具有显著的体内抗肿瘤活性。我们报道了细胞内药物浓度与7-OMen和阿霉素对培养的中国仓鼠卵巢(CHO)、B16和L1210细胞的杀伤作用之间存在的复杂关系。CHO、B16和L1210细胞对7-OMEN和阿霉素的摄取与细胞外药物浓度(微克/毫升)成正比。在相似浓度下，CHO和B16细胞比阿霉素积累更多的7种征兆。根据CHO细胞内的浓度，阿霉素[致死剂量为90%的细胞(LD90)，0.02微克/106细胞]比7-先兆(LD90，0.1微克/106细胞)更致命。然而，根据CHO细胞中的浓度，细胞外7-先兆的致死率(LD90，0.6微克/毫升)比阿霉素(LD90，2.5微克/毫升)更高。这可能是由于CHO细胞对阿霉素的摄取水平非常低。对于7-OMAN和阿霉素，细胞杀伤率均随细胞内浓度的增加而增加。与阿霉素的细胞内浓度与细胞杀伤率之间的线性关系相反，7-先兆的细胞内浓度与细胞杀伤率呈双相关系。对于7-征兆，细胞杀伤率与细胞内浓度成正比，最高可达0.04微克/106个细胞。进一步增加细胞内浓度并不会导致细胞杀伤率的成比例增加。指数生长的细胞不仅比平台期细胞积累了更多的7种征兆，而且天生对药物更敏感。后一个结论是基于观察到的，即使在相似的细胞内浓度，比平台期细胞更指数增长的细胞被杀死。荧光显微镜显示，7-征兆主要聚集在细胞质中，而阿霉素则聚集在细胞核中。讨论了这一观察结果对这些药物作用模式的意义。
Abstract 7-con-O-Methylnogarol (7-OMEN) is a new anthracycline antitumor agent with significant activity in vivo against murine P388 and L1210 leukemia (140% increase in life span) and B16 melanoma (114% increase in life span). We report here the complex relationship that exists between intracellular drug concentration and killing of Chinese hamster ovary (CHO), B16, and L1210 cells in culture by 7-OMEN and Adriamycin. The uptake of 7-OMEN and Adriamycin by CHO, B16, and L1210 cells was proportional to the extracellular concentration (µg/ml medium) of the drug. At similar concentrations, CHO and B16 cells accumulated more 7-OMEN than Adriamycin. Based on intracellular concentration in CHO cells, Adriamycin was more lethal [lethal dose for 90% of the cells (LD90), 0.02 µg/106 cells] than was 7-OMEN (LD90, 0.1 µg/106 cells). However, based on concentration in CHO cells extracellular 7-OMEN was more lethal (LD90, 0.6 µg/ml) than was Adriamycin (LD90, 2.5 µg/ml). This was probably due to the very low level of uptake of Adriamycin by CHO cells. For both 7-OMEN and Adriamycin, cell kill increased with increasing intracellular concentration. In contrast to the linear relationship between intracellular concentration and cell kill with Adriamycin, a biphasic relationship was seen with 7-OMEN. For 7-OMEN, cell kill was proportional to intracellular concentration up to approximately 0.04 µg/106 cells. Further increase in intracellular concentration did not result in proportional increase in cell kill. Exponentially growing cells not only accumulated more 7-OMEN than did plateau-phase cells but were also inherently more sensitive to the drug. This latter conclusion was based on the observation that, even at similar intracellular concentration, more exponentially growing than plateau-phase cells were killed. Fluorescence microscopy showed that 7-OMEN accumulated predominantly in the cytoplasm, whereas Adriamycin accumulated in the nucleus. The significance of this observation to the mode of action of these drugs is discussed.