Functional analysis of single nucleotide polymorphisms of hepatic organic anion transporter OATP1B1 (OATP-C)
Functional analysis of single nucleotide polymorphisms of hepatic organic anion transporter OATP1B1 (OATP-C)
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DOI：
10.1097/00008571-200411000-00006
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发表时间：
2004-11-01
期刊：
影响因子：
--
通讯作者：
Sugiyama, Y
中科院分区：
文献类型：
--
作者：
Iwai, M;Suzuki, H;Sugiyama, Y
目的在肝特异性转运蛋白--有机阴离子转运多肽1B 1（OATP 1B 1/OATP-C）基因上发现两种常见的单核苷酸多态性（SNP）：Asn 130 Asp和Val 174 Ala。虽然这两个SNPs在欧美人中独立存在，但在日本人中Val 174 Ala主要与Asn 130 Asp相关。我们之前在日本受试者中的体内研究表明，普伐他汀的非肾清除率降低至野生型受试者的13%（Nishizato et al. Clin Pharmacol Ther 2003;73（6）：554-564）。方法在HEK 293细胞中稳定表达野生型OATP 1B 1（OATP 1B 1 *1a）、OATP 1B 1 *1b（Asn 130 Asp）、OATP 1B 1 *5（Val 174 Ala）和OATP 1B 1 *15（Asn 130 Asp和Val 174 Ala），分析其定位和转运活性。为了表征固有的V-max，在摄取研究中观察到的V-max由Western blotting估计的表达水平标准化。在这些SNP变体中，未观察到17 β-雌二醇17 β-D-葡糖苷酸（E（2）17 β G）（OATP 1B 1的典型底物）转运的Km值发生显著变化。然而，与OATP 1B 1 *1a相比，OATP 1B 1 *15的归一化Vmax值大幅降低至小于30%。而OATP 1B 1 *1b（Asn 130 Asp）和OATP 1B 1 *5（Val 174 Ala）的转运活性与OATP 1B 1 * 1a相似。因此，它表明，在体内处置可以预测使用重组转运蛋白的体外结果。(C)2004年利平科特威廉姆斯威尔金斯。
Objective Two kinds of single nucleotide polymorphism (SNP; Asn130Asp and Val174Ala) are frequently observed in the liver specific transporter, organic anion transporting polypeptide 1B1 (OATP1B1/OATP-C) gene. Although these two SNPs occur independently in European-Americans, Val174Ala is mostly associated with Asn130Asp in Japanese. Our previous in-vivo studies in Japanese subjects indicated that the non-renal clearance of pravastatin was decreased to 13% of that in wild-type subjects (Nishizato et al. Clin Pharmacol Ther 2003;73(6):554-564). The purpose of the present study is to characterize the function of SNPs variants of OATP1B1 in cDNA transfected cells.Methods The localization and transport activity were analyzed in HEK293 cells stably expressing wild-type OATP1B1 (OATP1B1*1a), OATP1B1*1b (Asn130Asp), OATP1B1*5 (Val174Ala) and OATP1B1*15 (Asn130Asp and Val174Ala). To characterize the intrinsic V-max, observed V-max in uptake study were normalized by the expression level estimated from Western blotting.Results All SNP variants are predominantly located on the cell surface. No significant alteration was observed in Km values for the transport of 17beta-estradiol 17beta-D-glucuronide (E(2)17betaG), a typical substrate of OATP1B1, among these SNP variants. However, the normalized V-max value for OATP1B1*15 was drastically decreased to less than 30% compared with OATP1B1*1a. In contrast, the transport activity of OATP1B1*1b (Asn130Asp) and OATP1B1*5 (Val174Ala) was similar to that of OATP1B1*1a.Conclusions These results are consistent with the results of our previous clinical studies. It is thus suggested that in-vivo disposition may be predicted from in-vitro results using recombinant transporters. (C) 2004 Lippincott Williams Wilkins.