Rybczynska M, Liu R, Lu P, Sharom F J, Steinfels E, Pietro A D, Spitaler M, Grunicke H, Hofmann J
Institute of Medical Chemistry and Biochemistry, University of Innsbruck, Fritz-Pregl-Str. 3, Innsbruck, A-6020, Austria.
Br J Cancer. 2001 May 18;84(10):1405-11. doi: 10.1054/bjoc.2001.1776.
Miltefosine (hexadecylphosphocholine) is used for topical treatment of breast cancers. It has been shown previously that a high percentage of breast carcinomas express MDR1 or MRP. We investigated the sensitivity of MDR1 -expressing cells to treatment with miltefosine. We show that cells overexpressing MDR1 (NCI/ADR-RES, KB-8-5, KB-C1, CCRF/VCR1000, CCRF/ADR5000) were less sensitive to miltefosine treatment when compared to the sensitive parental cell lines. HeLa cells transfected with MDR1 exhibited resistance to the compound, indicating that expression of this gene is sufficient to reduce the sensitivity to miltefosine. The resistance of MDR1-expressing cells to miltefosine was less pronounced than that to adriamycin or vinblastine. Expression of MDR2 did not correlate with the resistance to miltefosine. As shown by a fluorescence quenching assay using MIANS-labelled P-glycoprotein (PGP), miltefosine bound to PGP with a K(d)of approximately 7 microM and inhibited PGP-ATPase activity with an IC(50)of approximately 35 microM. Verapamil was not able to reverse the resistance to miltefosine. Concentrations of miltefosine up to approximately 60 microM stimulated, whereas higher concentrations inhibited the transport of [3H]-colchicine with an IC(50)of approximately 297 microM. Binding studies indicated that miltefosine seems to interact with the transmembrane domain and not the cytosolic nucleotide-binding domain of PGP. These data indicate that expression of MDR1 may reduce the response to miltefosine in patients and that this compound interacts with PGP in a manner different from a number of other substrates.
米替福新(十六烷基磷酰胆碱)用于乳腺癌的局部治疗。先前已表明,高比例的乳腺癌表达多药耐药蛋白1(MDR1)或多药耐药相关蛋白(MRP)。我们研究了表达MDR1的细胞对米替福新治疗的敏感性。我们发现,与敏感的亲本细胞系相比,过表达MDR1的细胞(NCI/ADR-RES、KB-8-5、KB-C1、CCRF/VCR1000、CCRF/ADR5000)对米替福新治疗的敏感性较低。用MDR1转染的HeLa细胞对该化合物表现出抗性,表明该基因的表达足以降低对米替福新的敏感性。表达MDR1的细胞对米替福新的抗性不如对阿霉素或长春碱的抗性明显。MDR2的表达与对米替福新的抗性无关。如使用MIANS标记的P-糖蛋白(PGP)的荧光猝灭试验所示,米替福新与PGP结合,解离常数(K(d))约为7微摩尔,抑制PGP-ATP酶活性的半数抑制浓度(IC(50))约为35微摩尔。维拉帕米无法逆转对米替福新的抗性。高达约60微摩尔的米替福新浓度刺激了[3H]-秋水仙碱的转运,而更高浓度则抑制了其转运,半数抑制浓度(IC(50))约为297微摩尔。结合研究表明,米替福新似乎与PGP的跨膜结构域相互作用,而不是与胞质核苷酸结合结构域相互作用。这些数据表明,MDR1的表达可能降低患者对米替福新的反应,并且该化合物与PGP的相互作用方式与许多其他底物不同。