Characterization and analyses of multidrug resistance-associated protein 1 (MRP1/ABCC1) polymorphisms in Chinese population.
Characterization and analyses of multidrug resistance-associated protein 1 (MRP1/ABCC1) polymorphisms in Chinese population.
复制标题
中国人群多药耐药相关蛋白1（MRP1/ABCC1）多态性特征及分析
DOI：
10.1097/fpc.0b013e328323f680
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发表时间：
2009-03
影响因子：
2.6
通讯作者：
Liu ZQ
中科院分区：
文献类型：
--
作者：
Yin JY;Huang Q;Yang Y;Zhang JT;Zhong MZ;Zhou HH;Liu ZQ
目的探讨MRP 1/ABCC 1基因4种常见单核苷酸多态性（SNPs）在中国大陆人群中的分布频率，并探讨这些SNPs是否影响MRP 1/ABCC 1基因的表达和功能。方法采用聚合酶链反应-限制性片段长度多态性技术对208例健康志愿者进行基因分型。通过定点突变重新创建了这四个候选SNP，并在稳定转染的HEK 293和CHO-K1细胞系中检测了它们对MRP 1/ABCC 1表达和多药耐药功能的影响。采用实时荧光定量PCR、蛋白质印迹和共聚焦显微镜检测mRNA、蛋白质表达和蛋白质转运。最后，采用甲基噻唑四氮唑法检测突变对MRP 1/ABCC 1介导的耐药性的影响。结果Cys 43 Ser（128 G>C）、Thr 73 Ile（218 C>T）、Arg 723 Gln（2168 G>A）和Arg 1058 Gln（3173 G>A）等位基因频率在中国大陆人群中分别为0.5%、1.4%、5.8%和0.5%。这些突变对MRP 1/ABCC 1的表达和运输没有任何影响，但是Arg 723 Gln突变显著降低了MRP 1/ABCC 1介导的对柔红霉素、多柔比星、依托泊苷、长春碱和长春新碱的耐药性。Cys 43 Ser突变不影响所有测试的耐药性。相反，Thr 73 Ile突变降低了对甲氨蝶呤和依托泊苷的耐药性，而Arg 1058 Gln突变增加了HEK 293和CHO-K1细胞中两种蒽环类药物和依托泊苷以及CHO-K1细胞中长春碱和甲氨蝶呤的反应。结论Arg 723 Gln突变在中国大陆人群中的等位基因频率相对较高，该突变可显著降低MRP 1/ABCC 1在多药耐药中的活性。
Objective To explore the distribution frequencies of four common single nucleotide polymorphisms (SNPs) of MRP1/ABCC1 in a mainland Chinese population and investigate whether these SNPs affect the expression and function of the MRP1/ABCC1. Methods The genotype of 208 healthy volunteers was determined using PCR-restriction fragment length polymorphism. The four candidated SNPs were recreated by site-directed mutagenesis and tested for their effect on MRP1/ABCC1 expression and multidrug resistance function in stable transfected HEK293 and CHO-K1 cell lines. Real-time PCR, western blot and confocal microscopy were used to determine the mRNA, protein expression, and protein trafficking. At last, the effect of mutations on MRP1/ABCC1-mediate drug resistance was determined using methyl thiazolyl tetrazolium assay. Results The allelic frequencies of Cys43Ser (128G>C), Thr73Ile (218C>T), Arg723Gln (2168G>A), and Arg1058Gln (3173G>A) in mainland Chinese were 0.5, 1.4, 5.8, and 0.5%, respectively. None of these mutations had any effect on MRP1/ABCC1 expression and trafficking, but that Arg723Gln mutation significantly reduced MRP1/ABCC1-mediated resistance to daunorubicin, doxorubicin, etoposide, vinblastine, and vincristine. The Cys43Ser mutation did not affect all tested drug resistance. In contrast, the Thr73Ile mutation reduced resistance to methotrexate and etoposide, whereas the Arg1058Gln mutation increased the response of two anthracycline drugs and etoposide in HEK293 and CHO-K1 cells as well as vinblastine and methotrexate in CHO-K1 cells. Conclusion The allelic frequency of the Arg723Gln mutation is relatively higher than other SNPs in mainland Chinese population and therefore this mutation significantly reduces MRP1/ABCC1 activity in multidrug resistance.