A cell model for high-throughput screening GLP-1 receptor agonists

Rui WANG; Zi-Xin YANG; Zhi-Qiang KE; Zheng-Ding SU; Zhi-Qiang KE; Xi-Yao CHENG

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A cell model for high-throughput screening GLP-1 receptor agonists

Author: Rui WANG ¹ ; Zi-Xin YANG ¹ ; Zhi-Qiang KE ¹ ; Zheng-Ding SU ¹ ; Zhi-Qiang KE ² ; Xi-Yao CHENG ³
Author Information

1. Cooperative Innovation Center of Industrial Fermentation (Ministry of Education & Hubei Province), Hubei University of Technology
2. Hubei Key Laboratory of Diabetes and Angiopathy, Xianning Medical College, Hubei University of Science and Technology
3. School of Light Industry and Food Engineering, Guangxi University
Publication Type:Journal Article
Keywords: cAMP response element; GLP-1 receptor agonists; glucagon-like peptide-1; luciferase reporter gene; small-molecule GLP-1 receptor agonists; type 2 diabetes mellitus; Z' factor
From: Chinese Pharmacological Bulletin 2023;39(9):1793-1798
CountryChina
Language:Chinese
Abstract: Aim To establish a high-throughput screening cell model for GLP-1 receptor agonists. Methods A pEGFP-GLP-1R-3 C recombinant plasmid was constructed and transfected into HEK293T cells. The cells were screened with G418 and flow cytometry. The established stable cell line was named HEK293TGLP-lR-3C-eGFP cell line. The expression level of GLP-1 R-3C-eGFP protein was confirmed by Western blotting and laser confocal microscopy. Then cyclic adenosine monophosphate (cAMP) response element reporter gene was transfected into the HEK293T-GLP-lR-3C-eGFP cells. The luminescence values were detected by One-Step Luciferase Reporter Gene Assay Kit after stimulation with different concentrations of GLP-1 peptide. The luminescence values reflected the cellular cAMP level, which was verified using the cAMP kit (E L I S A). Results HEK293T-GLP-lR-3C-eGFP cell line was successfully constructed. The relative light unit change trend after stimulation with different concentrations of GLP-1 was similar to that of the cellular cAMP level change trend. The value of Z' in this experiment was 0.52. Conclusions A recombinant HEK293T cell line is established, which can be used for high-throughput screening of GLP-1 receptor agonists.