Abstract：Abstract: The human urate anion transporter (hURAT1) is thought to be an essential membrane protein that mediates the reabsorption of urate on the apical side of the proximal tubule. Screening hUTAT1 inhibitors has become a hot research topic in recent years. Therefore, construction of an in vitro hURAT1 inhibitor screening model is of significance in developing new uricosuric agents. Here, lentivirus vectors carrying SLC22A12 (hURAT1 coding gene) were constructed and then MDCK cells stably expressing hURAT1 (MDCK-URAT1) were established by lentivirus infection and G418 selection. The expression of hURAT1 in MDCK-URAT1 was validated by Western-blot and immunofluorescence. In addition, the [14C] uric acid uptake experiments were performed and the interactions of hURAT1 with benzbromarone and probenecid were examined. According to the results, a significant increase in not only hURAT1 expression but also uric acid uptake (with a Km value of 365.4 μmol/L) was shown in MDCK-URAT1(P>0.001). The IC50 values of the inhibition by benzbromarone and probenecid were 0.18 μmol/L and 66.82 μmol/L, respectively. In conclusion, a method for detecting the activity of hURAT1-targeting drugs in vitro was successfully established and would contribute to the development of uricosuric drugs.
引用本文:
陈嘉盛, 吴 婷, 丘玉昌, 曹 莹, 庞建新. 以hURAT1为靶点的排尿酸药物体外细胞筛选模型的建立和应用[J]. 生命科学研究, 2016, 20(3): 248-254. CHEN Jia-Sheng, WU Ting, QIU Yu-Chang, CAO Ying, PANG Jian-Xin. A Cell Line Stably Expressing Lentivirus-mediated hURAT1 as an In Vitro Model of Screening Uricosuric Agents. Life Science Research, 2016, 20(3): 248-254.
2016,Vol. 20
