Neubauer A, Valet G, Huhn D
Universitätsklinikum Rudolf-Virchow, Abteilung Innere Medizin, Berlin, FRG.
Anal Cell Pathol. 1989 Dec;2(1):49-58.
A recently developed flow cytometric assay method using patient tumor cells allows the determination not only of their sensitivity to cytostatic drugs but also of biochemical and biophysical parameters after treatment, such as esterase concentration and intracellular pH of the living cells. DNA-content of the dead cells and cell volume of living and dead cells. The T-cell lines CEM, Molt4, Jurkat, the B-cell lines RPMI1788, Daudi, Raji and the promyelocytic line HL60 were incubated with: cytosine arabinoside (ara-C), L-asparaginase, daunorubicin, vincristine and prednisone for 48 h. Living cells then stained with esterase and pH-dye 1,4-diacetoxy-2,3-dicyanobenzene (ADB) and dead cells with DNA-dye propidium-iodide (PI). The esterase concentration, an index of metabolic activity, decreased in the T-cell lines under the influence of ara-C, daunorubicin and vincristine, whereas in the B-cell lines smaller changes in esterase concentration were observed (P less than 0.001). A decrease in intracellular pH was seen in the ara-C and daunorubicin-incubated cells Molt4, CEM and HL60, whereas in the B-cell lines no significant change in intracellular pH was found. In all lines except Jurkat the cell volume of the surviving cells increased under the influence of certain drugs (primarily ara-C and daunorubicin); B-cell lines showed a greater swelling than T-cell lines (P = 0.001).
一种最近开发的使用患者肿瘤细胞的流式细胞术检测方法，不仅可以确定肿瘤细胞对细胞生长抑制剂的敏感性，还能测定治疗后细胞的生化和生物物理参数，如活细胞的酯酶浓度和细胞内pH值、死细胞的DNA含量以及活细胞和死细胞的细胞体积。将T细胞系CEM、Molt4、Jurkat，B细胞系RPMI1788、Daudi、Raji以及早幼粒细胞系HL60与阿糖胞苷（ara-C）、L-天冬酰胺酶、柔红霉素、长春新碱和泼尼松一起孵育48小时。然后用酯酶和pH染料1,4-二乙酰氧基-2,3-二氰基苯（ADB）对活细胞进行染色，用DNA染料碘化丙啶（PI）对死细胞进行染色。酯酶浓度是代谢活性的一个指标，在阿糖胞苷、柔红霉素和长春新碱的影响下，T细胞系中的酯酶浓度降低，而在B细胞系中，酯酶浓度变化较小（P<0.001）。在经阿糖胞苷和柔红霉素孵育的细胞Molt4、CEM和HL60中观察到细胞内pH值下降，而在B细胞系中未发现细胞内pH值有显著变化。除Jurkat外，在某些药物（主要是阿糖胞苷和柔红霉素）的影响下，所有细胞系中存活细胞的细胞体积均增加；B细胞系的肿胀程度大于T细胞系（P = 0.001）。
