Abstract：Abstract: To screen ATP binding cassette subfamily G member 2 (ABCG2) inhibitors, a stable ABCG2 expres-sion cell model was constructed. The expression vector pcDNA3.1(+)-ABCG2 was transfected into HEK293 cells using lipo3000, and the positive clones were selected by G418. The mRNA and protein expression le-vels in stable ABCG2 transgenic cells were validated by qRT-PCR and Western-blot methods. Membrane vesicles were then extracted by differential centrifugation and potential ABCG2 inhibitors were screened through 14C-uric acid uptake assay. The results showed that the mRNA and protein levels in ABCG2-ex-pressing cells were 1 717.5 and 2.64 times as much as those in mock-transfected cells, respectively. The uric acid uptake activity of the extracted vesicles from ABCG2-expressing cells was 3.73 times as much as that of control vesicles. In addition, the inhibitory effects of commonly used uric acid-lowering drugs on ABCG2 were evaluated using the obtained cell model. The results showed that benzbromarone inhibited ABCG2 with an IC50 value of (3.45±1.09) μmol/L, and RDEA3170 with an IC50 value of (9.90±2.11) μmol/L. In conclusion, the experiments showed successful construction of the screening model of ABCG2 inhibitors and also demon-strated for the first time the potential ABCG2 inhibition capability of RDEA3170.
引用本文:
郑凤欣, 赵泽安, 林雪曼, 吴 婷, 庞建新. 稳定表达ABCG2细胞模型的建立及其在筛选ABCG2抑制剂中的应用[J]. 生命科学研究, 2022, 26(2): 117-124.
ZHENG Feng-xin, ZHAO Ze-an, LIN Xue-man, WU Ting, PANG Jian-xin. Establishment of a Cell Model Stably Expressing ABCG2 and Its Application in Screening Inhibitors of ABCG2 . Life Science Research, 2022, 26(2): 117-124.
