Abstract
βKlotho as the major role is a necessary auxiliary protein when fibroblast growth factor 21 (FGF21) binds FGF21 receptors (FGFR) for activating intracellular signaling pathways that ultimately generate biological effects. To achieve the aim of high throughput screening of FGF21 analogues, we established 3T3-L1-βKlotho cells that could stably express βklotho protein. The glucose uptake, expression of GLUT1 mRNA and activation of FGF signaling molecules ERK1/2 phosphorylation were detected by GOD-POD assay, real-time PCR analysis and western blotting assay in 3T3-L1-βKlotho cells and 3T3-L1 adipocytes, respectively. The results showed that FGF21 increased glucose uptake significantly in a dose-dependent and time-dependent manner in 3T3-L1-βKlotho cells. 3T3-L1-βKlotho cells stimulated with FGF21 up-regulated the transcriptional levels of GLUT1 mRNA obviously. FGF21 activated the FGF signaling molecules ERK1/2 in 3T3-L1-βKlotho cells. In addition, the same results were obtained in 3T3-L1 adipocytes. Furthermore, FGF21-stimulated elevation of glucose uptake, GLUT1 mRNA transcription and the phosphorylation of ERK1/2 were dramatically attenuated by pretreatment of cells with FGFR specific inhibitor SU5402 in 3T3-L1-βKlotho cells. This study demonstrated that the cell model could be applied to high throughput screen FGF21 analogues.
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The research was financially supported by National Key R&D Program of China (2017YFD0501102, 2017YFD0501004), Science and Technology Planning Program of Heilongjiang Province (GC13C104), Research Fund for the Postdoctoral Program of Heilongjiang Province (LBH-Q09162), National Natural Science Foundation of China (J1210069/J0116).
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Guo, X., Wang, X., Yuan, Q. et al. Evaluation of a cell model expressing βKlotho for screening FGF21 analogues. Cytotechnology 71, 1033–1041 (2019). https://doi.org/10.1007/s10616-019-00344-z
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DOI: https://doi.org/10.1007/s10616-019-00344-z