Quality control of multidrug resistance assays in adult acute leukemia: Correlation between assays for P-glycoprotein expression and activity
Quality control of multidrug resistance assays in adult acute leukemia: Correlation between assays for P-glycoprotein expression and activity
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DOI：
10.1182/blood.v87.11.4809.bloodjournal87114809
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发表时间：
1996-06-01
期刊：
影响因子：
20.3
通讯作者：
Schuurhuis, GJ
中科院分区：
文献类型：
--
作者：
Broxterman, HJ;Sonneveld, P;Schuurhuis, GJ
我们比较了新鲜和解冻成人急性白血病中 P-糖蛋白 (Pgp/MDR1) 表型的多种测定，以验证和定量 Pgp 表达和功能的测量。结果与表达 Pgp 的 KB8 和 KB8-5 细胞系相关。最灵敏的测定是测量 2 μmol/L PSC833 对罗丹明 123 (R123) 荧光的调节。接下来是探针钙黄绿素-AM 的调制。我们还发现新鲜和解冻样品的 R123/PSC833 测定值之间存在良好的实验室内和实验室间相关性。此外，PSC833 对 H-3-柔红霉素 (DNR) 积累、DNR 荧光和 H-3-长春新碱积累的影响非常相似。 DNR/PSC833 和 R123/PSC833 测试之间的相关性为 r = 0.86 (N = 51)。 8 μmol/L 维拉帕米对药物蓄积的调节与 DNR 的 PSC833 效应相同（117%，N = 21），但在每个病例中长春新碱的效应更高（161% vs 121%，N = 22；P < .001），表明维拉帕米有额外的效应，与 Pgp 无关。 Pgp 活细胞染色与单克隆抗体 MRK16 的相关性对于 R123/PSC833 功能测试为 r = .77 (N = 52)，对于 DNR/PSC833 测试为 r = .84 (N = 50)。从这些结果可以计算出，通过在具有最高平均 Pgp 表达的白血病原始细胞样品中用 PSC833 阻断 Pgp 泵活性，可以实现平均 DNR 积累的最大增加约 50%。可能存在具有较高 Pgp 活性的母细胞亚群。它们的相关性必须进一步研究。本文概述的方法允许在多中心临床 Pgp 调制研究中对白血病中的 Pgp/MDR1 表型进行可靠、定量监测。 (C) 1996 年，美国血液学会。
We have compared multiple assays for the P-glycoprotein (Pgp/MDR1) phenotype in fresh and thawed adult acute leukemia to validate and quantitate measures for the expression and function of Pgp. The results are related to the Pgp-expressing KB8 and KB8-5 cell lines. The most sensitive assay was the measurement of modulation of the rhodamine 123 (R123) fluorescence by 2 mu mol/L PSC833. followed by the modulation of the probe calcein-AM. We also found a good intralaboratory and interlaboratory correlation between the values of the R123/PSC833 assay for fresh as well as thawed samples. In addition, the effects of PSC833 on H-3-daunorubicin (DNR) accumulation, DNR fluorescence, and H-3-vincristine accumulation were very similar. The correlation between the DNR/PSC833 and R123/PSC833 test was r = .86 (N = 51). The modulation of drug accumulation by 8 mu mol/L verapamil was the same as the PSC833 effect for DNR (117%, N = 21), but was higher for vincristine in every single case (161% v 121%, N = 22; P < .001), indicating additional verapamil effects, not related to Pgp. The correlation of the staining of viable cells for Pgp with the monoclonal antibody MRK16 was r = .77 (N = 52) for the R123/PSC833 functional test and r = .84 (N = 50) for the DNR/PSC833 test. From these results it could be calculated that a maximal increase of the mean DNR accumulation of about 50% can be achieved by blocking Pgp pump activity with PSC833 in leukemic blast samples with the highest mean Pgp expression. Subpopulations of blast cells with higher Pgp activity are likely to be present. Their relevance has to be studied further. The methods outlined here allow the reliable, quantitative monitoring of the Pgp/MDR1 phenotype in leukemias in multicentered, clinical Pgp modulation studies. (C) 1996 by The American Society of Hematology.