Department of Clinical Pharmacology and Pharmacoepidemiology, Medical Faculty Heidelberg, Heidelberg University Hospital, Heidelberg University, Im Neuenheimer Feld 410, 69120 Heidelberg, Germany.
Department of Food and Drug, University of Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy.
Int J Mol Sci. 2024 Apr 8;25(7):4118. doi: 10.3390/ijms25074118.
Drug efflux transporters of the ATP-binding-cassette superfamily play a major role in the availability and concentration of drugs at their site of action. ABCC2 (MRP2) and ABCG2 (BCRP) are among the most important drug transporters that determine the pharmacokinetics of many drugs and whose overexpression is associated with cancer chemoresistance. ABCC2 and ABCG2 expression is frequently altered during treatment, thus influencing efficacy and toxicity. Currently, there are no routine approaches available to closely monitor transporter expression. Here, we developed and validated a UPLC-MS/MS method to quantify ABCC2 and ABCG2 in extracellular vesicles (EVs) from cell culture and plasma. In this way, an association between ABCC2 protein levels and transporter activity in HepG2 cells treated with rifampicin and hypericin and their derived EVs was observed. Although ABCG2 was detected in MCF7 cell-derived EVs, the transporter levels in the vesicles did not reflect the expression in the cells. An analysis of plasma EVs from healthy volunteers confirmed, for the first time at the protein level, the presence of both transporters in more than half of the samples. Our findings support the potential of analyzing ABC transporters, and especially ABCC2, in EVs to estimate the transporter expression in HepG2 cells.
三磷酸腺苷结合盒超家族的药物外排转运蛋白在药物在作用部位的可用性和浓度方面发挥着重要作用。ABCC2（MRP2）和 ABCG2（BCRP）是最重要的药物转运蛋白之一，它们决定了许多药物的药代动力学，其过度表达与癌症化疗耐药性有关。ABCC2 和 ABCG2 的表达在治疗过程中经常发生改变，从而影响疗效和毒性。目前，尚无常规方法可用于密切监测转运蛋白的表达。在这里，我们开发并验证了一种用于定量细胞培养和血浆中外泌体（EVs）中 ABCC2 和 ABCG2 的 UPLC-MS/MS 方法。通过这种方式，观察到在用利福平（rifampicin）和金丝桃素（hypericin）处理的 HepG2 细胞及其衍生的 EVs 中，ABCC2 蛋白水平与转运体活性之间存在关联。尽管在 MCF7 细胞衍生的 EVs 中检测到了 ABCG2，但囊泡中的转运体水平并未反映出细胞中的表达水平。对来自健康志愿者的血浆 EVs 的分析首次在蛋白质水平上证实了这两种转运体在超过一半的样本中的存在。我们的研究结果支持分析 EVs 中的 ABC 转运蛋白，特别是 ABCC2，以估计 HepG2 细胞中转运体表达的潜力。
