Clinical Pharmacology, Division of Drug Research, Department of Medical and Health Sciences, Linköping University, Linköping, Sweden.
Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden.
Pharmacol Res Perspect. 2020 Apr;8(2):e00581. doi: 10.1002/prp2.581.
Single nucleotide polymorphisms (SNPs) in the gene for multidrug resistance protein ABCG2, an erlotinib transporter, is a possible contributor to the interindividual variation observed in erlotinib pharmacokinetics and toxicity. Therefore, the aim was to study erlotinib efflux by ABCG2 wild-type (wt) and ABCG2 polymorphic variants in the K562 cell line. The chronic myeloid leukemia K562 cell line, neither expressing EGFR nor ABCG2, was transduced with vectors containing the ABCG2 wt, the SNPs: 34 G > A and 421 C > A, or with empty vector (K562/ve). ABCG2-expressing cells were enriched using magnetic sorting and the expression was verified using flow cytometry. Intracellular erlotinib concentrations were analyzed by LC-MS/MS after incubation with 1 µmol/L erlotinib for 60 minutes. All recombinant cell lines were confirmed carriers of the vector and expressed ABCG2. Differences in intracellular erlotinib concentrations were observed between K562/ve and K562 ABCG2 wt and between K562/ve and K562 ABCG2 34G > A (both P ≤ .001, one-way ANOVA with Tukey HSD post hoc test), indicating that the cell lines carrying ABCG2 wt and ABCG2 34G > A actively transports erlotinib out of the cells. The ABCG2 34G > A cell line had a higher transport capacity compared with ABCG2 wt after adjusting for ABCG2 expression (P = .024, t test). No differences were observed between K562/ve and K562 ABCG2 421 C > A. Genetic polymorphism in the ABCG2 gene has an influence on the transport of erlotinib which can contribute to the observed variation in erlotinib pharmacokinetics and toxicity.
单核苷酸多态性(SNP)在多药耐药蛋白 ABCG2 基因中,ABCG2 是一种表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)的转运体,可能是导致厄洛替尼药代动力学和毒性个体间差异的原因之一。因此,本研究旨在探讨 ABCG2 野生型(wt)和 ABCG2 多态性变异体在 K562 细胞系中对厄洛替尼外排的影响。慢性髓系白血病 K562 细胞系既不表达 EGFR 也不表达 ABCG2,用含有 ABCG2wt、SNP:34G>A 和 421C>A 或空载载体(K562/ve)的载体转导。用磁珠分选法富集 ABCG2 表达细胞,并通过流式细胞术验证表达。用 LC-MS/MS 分析孵育 60 分钟后 1µmol/L 厄洛替尼时细胞内的厄洛替尼浓度。所有重组细胞系均证实携带载体并表达 ABCG2。K562/ve 与 K562 ABCG2wt 之间以及 K562/ve 与 K562 ABCG234G>A 之间(均 P≤.001,单因素方差分析加 Tukey HSD 事后检验)观察到细胞内厄洛替尼浓度存在差异,这表明携带 ABCG2wt 和 ABCG234G>A 的细胞系可主动将厄洛替尼从细胞内转运出去。在调整 ABCG2 表达后,ABCG234G>A 细胞系的转运能力高于 ABCG2wt(P=.024,t 检验)。K562/ve 与 K562 ABCG2421C>A 之间未见差异。ABCG2 基因的遗传多态性对厄洛替尼的转运有影响,这可能导致厄洛替尼药代动力学和毒性的个体间差异。