Overexpression of the multidrug resistance genes mdr1, mdr3, and mrp in L1210 leukemia cells resistant to inhibitors of ribonucleotide reductase
Overexpression of the multidrug resistance genes mdr1, mdr3, and mrp in L1210 leukemia cells resistant to inhibitors of ribonucleotide reductase
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DOI：
10.1016/s0006-2952(97)00210-4
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发表时间：
1997-09-15
影响因子：
5.8
通讯作者：
Sartorelli, AC
中科院分区：
文献类型：
--
作者：
Rappa, G;Lorico, A;Sartorelli, AC
L1210 MQ-580 是一种鼠白血病细胞系，对 α-(N)-杂环甲醛缩氨基硫脲类核糖核苷酸还原酶抑制剂的细胞毒性活性具有抵抗力。该品系对依托泊苷、道诺霉素和长春花碱具有交叉耐药性。 L1210 MQ-580 细胞对 3-氨基吡啶-2-甲醛缩氨基硫脲 (3-AP) 表现出 8 倍的耐药性，3-AP 是一种相对较新开发的核糖核苷酸还原酶抑制剂。 L1210 MQ-580 细胞的 [C-14]3-AP 积累量比亲代 L1210 细胞少 5 至 6 倍。 3-AP 流出速率增加是耐药细胞中 3-AP 稳态浓度较低的原因。在逆转录聚合酶链式反应测定中，与亲本 L1210 细胞相比，L1210 MQ-580 细胞过度表达多药耐药基因 mdr1、mdr3 和 mrp，但不过度表达 mdr2 基因。阿霉素（mdr 和 mrp 基因产物的潜在底物）的稳态浓度测量表明，L1210 MQ-580 细胞积累的蒽环类药物比亲代细胞少 4 倍。这些发现表明药物外流是 L1210 MQ-580 细胞交叉耐药模式的主要决定因素。为了将这些观察结果外推到 mdr1、mdr3 和 mrp 鼠基因的人类同源物，分别在用人 MDR1 和 MRP cDNA 转染的 L1210/VMDRC0.06 和 NIH3T3 36-8-32 细胞中测量 3-AP 的影响。转染子对 3-AP 的细胞毒性作用的抵抗力是其亲代模拟转染子的 2 至 3 倍，并且积累的 [C-14]3-AP 较少。此外，在两个双mrp基因敲除细胞系中，3-AP的细胞毒活性显着高于亲本W 9.5胚胎干细胞。因此，结果表明 3-AP 是 P-糖蛋白和 MRP 的底物，并且基线 MRP 表达能够发挥针对该药物毒性的保护作用。 (C) 1997 爱思唯尔科学公司。
L1210 MQ-580 is a murine leukemia cell line resistant to the cytotoxic activity of the alpha-(N)-heterocyclic carboxaldehyde thiosemicarbazone class of inhibitors of ribonucleotide reductase. The line is cross-resistant to etoposide, daunomycin, and vinblastine. L1210 MQ-580 cells expressed 8-fold resistance to 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP), a relatively newly developed inhibitor of ribonucleotide reductase. The accumulation of [C-14]3-AP by L1210 MQ-580 cells was 5- to 6-fold less than by parental L1210 cells. An increased rate of efflux of 3-AP was responsible for the lower steady-state concentration of 3-AP in resistant cells. In reverse transcription-polymerase chain reaction assays, L1210 MQ-580 cells were found to overexpress the multidrug resistance genes mdr1, mdr3, and mrp, but not the mdr2 gene, compared with parental L1210 cells. Measurement of the steady-state concentration of doxorubicin, a potential substrate for both the mdr and mrp gene products, demonstrated that L1210 MQ-580 cells accumulated 4-fold less anthracycline than parental cells. These findings indicate that drug efflux is a major determinant of the pattern of cross-resistance of L1210 MQ-580 cells. To extrapolate these observations to the human homologues of the mdr1, mdr3, and mrp murine genes, the effects of 3-AP were measured in L1210/VMDRC0.06 and NIH3T3 36-8-32 cells transfected with human MDR1 and MRP cDNAs, respectively. The transfectants were 2- to 3-fold resistant to the cytotoxic effects of 3-AP and accumulated less [C-14]3-AP than their parental mock-transfected counterparts. Moreover, the cytotoxic activity of 3-AP was significantly greater in two double mrp gene knockout cell lines than in parental W 9.5 embryonic stem cells. Thus, the results suggest that 3-AP is a substrate for both the P-glycoprotein and MRP and that baseline MRP expression has the capacity to exert a protective role against the toxicity of this agent. (C) 1997 Elsevier Science Inc.