Tatu C A, Paunescu V, Stanescu D I, Schneider F
Forslys Group, Timisoara, Romania. calin@umf-forslys sorostm.ro
Anticancer Res. 1997 Jul-Aug;17(4A):2583-6.
A relatively common and frequent form of multidrug resistance(MDR) in cancer cells is due to membrane overexpression of P-glycoprotein. Mdr phenotype was investigated by flow-cytometry in several types of malignant hemopathies -chronic lymphocytic leukemia, non-Hodgkin lymphomas, acute lymphoblastic and myeloblastic leukemias. We used daunomycin and fluo-3 as fluorochromes, and verapamil as reversor agent. The method is lacking unitary clinical parametrization and in order to improve it, we tried to establish an optimal concentration of verapamil, which was shown to be 14.92 micrograms/ml. The reliability of results obtained with fluo-3 in culture media containing Ca2+ is questionable, as low variations in the intracellular level of this ion dramatically influences light emission by the fluorochrome and possibly the function of P-gp. To avoid such fluorescence intensity variations, Ca(2+)-free cell culture medium for fluo-3-based flow-cytometric assay is suggested to be used.
癌细胞中一种相对常见且频繁出现的多药耐药（MDR）形式是由于P-糖蛋白的膜过度表达。通过流式细胞术在几种恶性血液病——慢性淋巴细胞白血病、非霍奇金淋巴瘤、急性淋巴细胞白血病和急性髓细胞白血病中研究了Mdr表型。我们使用柔红霉素和Fluo-3作为荧光染料，维拉帕米作为逆转剂。该方法缺乏统一的临床参数化，为了改进它，我们试图确定维拉帕米的最佳浓度，结果显示为14.92微克/毫升。在含有Ca2+的培养基中使用Fluo-3获得的结果的可靠性值得怀疑，因为该离子细胞内水平的微小变化会显著影响荧光染料的发光，并可能影响P-糖蛋白的功能。为避免此类荧光强度变化，建议使用基于Fluo-3的流式细胞术检测的无Ca(2+)细胞培养基。
