Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Radboudumc, Nijmegen, The Netherlands (T.T.G.N., J.G.P.P., D.D., D.K., K.J., A.H.V.A., F.G.M.R., M.J.W.); Department of Physical Chemistry/Bioenergetics, Institute of Chemistry PC14, Technical University of Berlin, Berlin, Germany (N.N.T., T.F.); and Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht, The Netherlands (K.J., R.M.).
Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Radboudumc, Nijmegen, The Netherlands (T.T.G.N., J.G.P.P., D.D., D.K., K.J., A.H.V.A., F.G.M.R., M.J.W.); Department of Physical Chemistry/Bioenergetics, Institute of Chemistry PC14, Technical University of Berlin, Berlin, Germany (N.N.T., T.F.); and Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht, The Netherlands (K.J., R.M.)
Drug Metab Dispos. 2018 May;46(5):592-599. doi: 10.1124/dmd.117.079384. Epub 2018 Mar 7.
Cisplatin is a cytostatic drug used for treatment of solid organ tumors. The main adverse effect is organic cation transporter 2 (OCT2)-mediated nephrotoxicity, observed in 30% of patients. The contribution of other renal drug transporters is elusive. Here, cisplatin-induced toxicity was evaluated in human-derived conditionally immortalized proximal tubule epithelial cells (ciPTEC) expressing renal drug transporters, including OCT2 and organic anion transporters 1 (OAT1) or 3 (OAT3). Parent ciPTEC demonstrated OCT2-dependent cisplatin toxicity (TC 34 ± 1 M after 24-hour exposure), as determined by cell viability. Overexpression of OAT1 and OAT3 resulted in reduced sensitivity to cisplatin (TC 45 ± 6 and 64 ± 11 M after 24-hour exposure, respectively). This effect was independent of OAT-mediated transport, as the OAT substrates probenecid and diclofenac did not influence cytotoxicity. Decreased cisplatin sensitivity in OAT-expressing cells was associated directly with a trend toward reduced intracellular cisplatin accumulation, explained by reduced OCT2 gene expression and activity. This was evaluated by V of the OCT2-model substrate ASP (23.5 ± 0.1, 13.1 ± 0.3, and 21.6 ± 0.6 minutes in ciPTEC-parent, ciPTEC-OAT1, and ciPTEC-OAT3, respectively). Although gene expression of cisplatin efflux transporter multidrug and toxin extrusion 1 (MATE1) was 16.2 ± 0.3-fold upregulated in ciPTEC-OAT1 and 6.1 ± 0.7-fold in ciPTEC-OAT3, toxicity was unaffected by the MATE substrate pyrimethamine, suggesting that MATE1 does not play a role in the current experimental set-up. In conclusion, OAT expression results in reduced cisplatin sensitivity in renal proximal tubule cells, explained by reduced OCT2-mediated uptake capacity. In vitro drug-induced toxicity studies should consider models that express both OCT and OAT drug transporters.
顺铂是一种细胞毒性药物，用于治疗实体器官肿瘤。其主要的不良反应是有机阳离子转运体 2（OCT2）介导的肾毒性，约 30%的患者会出现这种情况。其他肾脏药物转运体的作用尚不清楚。在这里，我们评估了在表达肾脏药物转运体（包括 OCT2 和有机阴离子转运体 1（OAT1）或 3（OAT3）的人源条件永生化近端肾小管上皮细胞（ciPTEC）中，顺铂诱导的毒性。亲本 ciPTEC 表现出 OCT2 依赖性顺铂毒性（暴露 24 小时后 TC 34±1μM），通过细胞活力测定。OAT1 和 OAT3 的过表达导致对顺铂的敏感性降低（暴露 24 小时后 TC 45±6 和 64±11μM）。这种作用与 OAT 介导的转运无关，因为 OAT 底物丙磺舒和双氯芬酸不影响细胞毒性。在表达 OAT 的细胞中，顺铂敏感性降低与细胞内顺铂积累减少直接相关，这可以用 OCT2 基因表达和活性降低来解释。这是通过评估 OCT2 模型底物 ASP 的 V 来评估的（在 ciPTEC-亲本、ciPTEC-OAT1 和 ciPTEC-OAT3 中分别为 23.5±0.1、13.1±0.3 和 21.6±0.6 分钟）。尽管 ciPTEC-OAT1 中的多药和毒素外排蛋白 1（MATE1）的基因表达上调了 16.2±0.3 倍，ciPTEC-OAT3 中上调了 6.1±0.7 倍，但嘧啶甲胺作为 MATE 底物对毒性没有影响，这表明 MATE1 在当前的实验设置中不起作用。总之，OAT 的表达导致肾脏近端肾小管细胞对顺铂的敏感性降低，这可以用 OCT2 介导的摄取能力降低来解释。在体外药物诱导的毒性研究中，应考虑表达 OCT 和 OAT 药物转运体的模型。
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