Legrand O, Simonin G, Perrot J Y, Zittoun R, Marie J P
Service d'Hématologie Clinique, the Service d'Hématologie Biologique, and the Formation de Recherche Associée Claude Bernard sur le Traitement des Hémopathies Malignes, Laboratoire Universitaire Paris VI, Paris, France.
Blood. 1998 Jun 15;91(12):4480-8.
Thirteen cell lines with different levels of Pgp and MRP expression were used to assess the ability of calcein acetoxymethyl ester (calcein-AM) uptake and calcein efflux to measure Pgp and MRP functions, respectively. There was a good correlation between MRP expression and the modulatory effect of probenecid (a specific modulator of MRP) on the calcein efflux (r = .91, P = .0003) and between Pgp expression and the modulatory effect of CsA on calcein-AM uptake (r = .96, P < .0001). In light of the high correlations for both proteins, we tested calcein-AM uptake and efflux in fresh myeloid leukemic cells. In 53 acute myeloid leukemia (AML) patients, there was also a good correlation between MRP expression (measured by reverse transcription-polymerase chain reaction and by MRPm6 expression by flow cytometry) and the modulatory effect of probenecid on the calcein fluorescence (r = .92, P < .0001) and between Pgp expression as measured by UIC2 antibody binding on flow cytometry and the modulatory effect of cyclosporin A on calcein-AM uptake (r = .83, P < .0001). Pgp activity was higher in CD34+ leukemia than in CD34- leukemia (2.26 +/- 1.50 v 1.46 +/- 1.21, respectively; P = .003), and MRP activity was higher in CD34- leukemia than in CD34+ leukemia (1.77 +/- 0.40 v 1.4 +/- 0. 29, respectively; P = .004). Pgp expression and activity (P = .004 and P = .01, respectively) and MRP activity (P = .03) but not MRP expression were prognostic factors for achievement of complete remission. These results suggest that functional testing (with calcein-AM +/- modulators) for the presence of both MRP and Pgp activities is of prognostic value and that MRP contributes to drug resistance in AML.
使用13种具有不同Pgp和MRP表达水平的细胞系来分别评估乙酰氧基甲基钙黄绿素（calcein-AM）摄取和钙黄绿素外排测量Pgp和MRP功能的能力。MRP表达与丙磺舒（MRP的一种特异性调节剂）对钙黄绿素外排的调节作用之间存在良好的相关性（r = 0.91，P = 0.0003），Pgp表达与环孢素A对calcein-AM摄取的调节作用之间也存在良好的相关性（r = 0.96，P < 0.0001）。鉴于两种蛋白的相关性都很高，我们在新鲜的髓系白血病细胞中测试了calcein-AM摄取和外排。在53例急性髓系白血病（AML）患者中，MRP表达（通过逆转录-聚合酶链反应和流式细胞术检测MRPm6表达来测量）与丙磺舒对钙黄绿素荧光的调节作用之间也存在良好的相关性（r = 0.92，P < 0.0001），通过流式细胞术检测UIC2抗体结合来测量的Pgp表达与环孢素A对calcein-AM摄取的调节作用之间也存在良好的相关性（r = 0.83，P < 0.0001）。CD34+白血病中的Pgp活性高于CD34-白血病（分别为2.26±1.50和1.46±1.21；P = 0.003），CD34-白血病中的MRP活性高于CD34+白血病（分别为1.77±0.40和1.4±0.29；P = 0.004）。Pgp表达和活性（分别为P = 0.004和P = 0.01）以及MRP活性（P = 0.03）而非MRP表达是实现完全缓解的预后因素。这些结果表明，针对MRP和Pgp活性的功能性检测（使用calcein-AM ± 调节剂）具有预后价值，并且MRP在AML的耐药性中起作用。
