阿帕替尼调控SOX5/GLUT4信号轴对胃癌细胞糖酵解的影响

Effects of apatinib regulating SOX5/GLUT4 signal axis on glycolysis of gastric cancer cells

目的探讨阿帕替尼在胃癌细胞糖酵解中的作用及机制。方法采用qRT-PCR和Western blotting评估SOX5和GLUT4的mRNA和蛋白在胃癌细胞中的表达,CCK8检测细胞活力。使用葡萄糖摄取比色测定试剂盒测定葡萄糖摄取,乳酸比色测定试剂盒对乳酸水平进行定量。细胞代谢分析仪检测各组细胞的细胞外酸化速率,并计算糖酵解能力值及糖酵解保留值。用双荧光素酶实验来评估GLUT4启动子活性。结果与对照组相比,阿帕替尼处理组胃癌细胞活力和糖酵解能力均下降。与oe-NC组相比,阿帕替尼处理后SOX5和GLUT4的表达量以及GLUT4启动子活性显著下调。当阿帕替尼处理的同时过表达SOX5,SOX5和GLUT4的表达量以及GLUT4启动子活性相比于仅用阿帕替尼处理显著上调。用阿帕替尼处理并同时过表达SOX5或GLUT4后细胞生存率和糖酵解能力相较于仅用阿帕替尼处理均发生显著上调。结论阿帕替尼通过抑制SOX5/GLUT4信号轴抑制胃癌细胞的细胞增殖和糖酵解。

Objective To clarify the role and mechanism of apatinib in glycolysis of gastric cancer cells.Methods qRT-PCR and western blotting were used to evaluate the mRNA and protein expression of SOX5 and GLUT4 in gastric cancer cells,and CCK8 was used to detect cell viability.Glucose uptake was measured using a glucose uptake colorimetric assay kit,and a lactic acid colorimetric assay kit was used to quantify the level of lactic acid.The cell metabolism analyzer was used to detect the extracellular acidification rate of each group of cells,and calculate the glycolysis capacity value and glycolysis retention value.A dual luciferase experiment was used to evaluate the GLUT4 promoter activity.Results Compared with the control group,the vitality and glycolytic ability of gastric cancer cells were decreased in the apatinib treatment group.Compared with the oe-NC group,the expression of SOX5 and GLUT4 and the activity of GLUT4 promoter were significantly down-regulated after apatinib treatment.When SOX5 was overexpressed while being treated with apatinib,the expression of SOX5 and GLUT4 and the activity of the GLUT4 promoter were significantly up-regulated compared with treatment with only apatinib.After treatment with apatinib and overexpression of SOX5 or GLUT4 at the same time,the cell survival rate and glycolytic capacity were significantly up-regulated compared with the treatment with apatinib alone.Conclusion Apatinib inhibits cell proliferation and glycolysis of gastric cancer cells by downregulation of the SOX5/GLUT4 signal axis.