Anderson Adrian B, Arriaga Edgar A
Department of Chemistry, University of Minnesota, 207 Pleasant Street SE, Minneapolis, MN 55455, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Sep 5;808(2):295-302. doi: 10.1016/j.jchromb.2004.05.017.
Micellar electrokinetic capillary chromatography with laser-induced fluorescence detection was used to detect the differences in doxorubicin metabolite accumulation in four subcellular fractions isolated from the CCRF-CEM and the CEM/C2 human leukemia cell lines. Five fluorescent metabolites and doxorubicin make up the metabolite profile of these cell lines upon treatment with 10 microM doxorubicin for 12 h, cell lysis, and fractionation by differential centrifugation. Based on the relative electrophoretic mobility of synthetic standards, we tentatively identify one metabolite as 7-deoxydoxorubicinone and suggest that doxorubicinone is not among those metabolites detected. Although the obvious difference between the derived cell line (CEM/C2) and the parent cell line (CCRF-CEM) is the decreased topoisomerase I activity in the former, the results presented here indicate that each cell line has a unique distribution of metabolites in each one of four subcellular fractions: nuclear-enriched, heavy-organelle-enriched, light-organelle-enriched, and cytoplasmic fractions.
采用胶束电动毛细管色谱法结合激光诱导荧光检测，以检测从CCRF - CEM和CEM/C2人白血病细胞系分离出的四个亚细胞组分中阿霉素代谢物积累的差异。在用10微摩尔阿霉素处理12小时、细胞裂解并通过差速离心分级分离后，五种荧光代谢物和阿霉素构成了这些细胞系的代谢物谱。基于合成标准品的相对电泳迁移率，我们初步鉴定出一种代谢物为7 - 脱氧阿霉素酮，并表明阿霉素酮不在检测到的那些代谢物之中。尽管衍生细胞系（CEM/C2）与亲本细胞系（CCRF - CEM）之间的明显差异在于前者拓扑异构酶I活性降低，但此处给出的结果表明，每个细胞系在四个亚细胞组分（富含细胞核的、富含重细胞器的、富含轻细胞器的和细胞质组分）中的每一个中都有独特的代谢物分布。
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