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.

中文翻译：

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评价表达βKlotho的细胞模型以筛选FGF21类似物。

当成纤维细胞生长因子21（FGF21）与FGF21受体（FGFR）结合以激活细胞内信号通路并最终产生生物学效应时，βKlotho是主要的必需蛋白。为了实现高通量筛选FGF21类似物的目的，我们建立了可以稳定表达βklotho蛋白的3T3-L1-βKlotho细胞。在3T3-L1-βKlotho细胞和3T3-L1脂肪细胞中分别通过GOD-POD测定，实时PCR分析和western印迹测定检测葡萄糖摄取，GLUT1 mRNA表达和FGF信号分子ERK1 / 2磷酸化激活。结果表明，FGF21在3T3-L1-βKlotho细胞中以剂量依赖和时间依赖的方式显着增加了葡萄糖的摄取。FGF21刺激的3T3-L1-βKlotho细胞明显上调GLUT1 mRNA的转录水平。FGF21激活3T3-L1-βKlotho细胞中的FGF信号分子ERK1 / 2。此外，在3T3-L1脂肪细胞中也获得了相同的结果。此外，通过在3T3-L1-βKlotho细胞中用FGFR特异性抑制剂SU5402预处理细胞，可显着减弱FGF21刺激的葡萄糖摄取升高，GLUT1 mRNA转录和ERK1 / 2磷酸化。这项研究表明，该细胞模型可以应用于高通量筛选FGF21类似物。通过在3T3-L1-βKlotho细胞中用FGFR特异性抑制剂SU5402预处理细胞，可显着减弱GLUT1 mRNA的转录和ERK1 / 2的磷酸化。这项研究表明，该细胞模型可以应用于高通量筛选FGF21类似物。通过在3T3-L1-βKlotho细胞中用FGFR特异性抑制剂SU5402预处理细胞，可显着减弱GLUT1 mRNA的转录和ERK1 / 2的磷酸化。这项研究表明该细胞模型可以应用于高通量筛选FGF21类似物。