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神经降压素受体-1(neurotensin receptor-1,NTR1)是G蛋白偶联受体家族的成员,调节细胞内钙离子的水平。NTR1与多种疾病的发生发展密切相关。为寻找与NTR1相关的拮抗剂,建立了稳定表达人源NTR1蛋白的中国仓鼠卵巢细胞系CHO/NTR1。将重组质粒利用脂质体转染的方法转入CHO细胞,加入G418抗生素进行筛选,采用Western blotting和钙离子浓度检测来评判CHO/NTR1细胞系是否构建成功。通过钙离子浓度检测,可以得到NTR1的天然配体神经降压素(neurotensin,NT)的EC50值和NTR1的已知拮抗剂SR48692的IC50值。结果提示,人源CHO/NTR1稳转细胞系可以用来研究NTR1的生物学特征和进一步筛选NTR1的拮抗剂与激动剂。
Neurotensin receptor-1 (NTR1) is a member of the G-protein coupled receptor family that regulates intracellular calcium levels. NTR1 and a variety of diseases are closely related to the occurrence and development. To find an antagonist related to NTR1, a Chinese hamster ovary cell line CHO / NTR1 stably expressing the human NTR1 protein was established. The recombinant plasmids were transfected into CHO cells by liposome transfection and G418 antibiotics were selected for screening. The construction of CHO / NTR1 cell lines was judged by Western blotting and calcium ion concentration test. The EC50 value of the natural ligand neurotensin (NT) of NTR1 and the IC50 value of the known antagonist SR48692 of NTR1 can be obtained by calcium ion concentration detection. The results suggest that human CHO / NTR1 stable cell line can be used to study the biological characteristics of NTR1 and further screening of antagonists and agonists of NTR1.