Background

Apatinib is a tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor-2 (VEGFR2), and has shown encouraging therapeutic effects in various malignant tumors. As yet, however, the role of apatinib in ovarian cancer has remained unknown. Here, we sought to elucidate the role of apatinib in the in vitro and in vivo viability and proliferation of ovarian cancer cells, as well as in glucose metabolism in these cells.

Methods

The effects of apatinib on ovarian cancer cell viability and proliferation were assessed using Cell Counting Kit-8 (CCK-8) and colony formation assays, respectively. The expression of VEGFR2/AKT1/SOX5/GLUT4 pathway proteins was assessed using Western blotting, and glucose uptake and lactate production assays were used to detect glycolysis in ovarian cancer cells. SOX5 was exogenously over-expressed and silenced in ovarian cancer cells using expression vector and shRNA-based methods, respectively. RNA expression analyses were performed using RNA-seq and gene-chip-based methods. GLUT4 promoter activity was assessed using a dual-luciferase reporter assay. The expression of p-VEGFR2 (Tyr1175), p-AKT1 (Ser473), p-GSK3β (Ser9), SOX5 and GLUT4 in xenograft tissues was assessed using immunohistochemistry (IHC).

Results

We found that apatinib inhibited the in vitro and in vivo viability and proliferation in Hey and OVCA433 ovarian cancer cells in a dose-dependent and time-dependent manner. We also found that apatinib effectively suppressed glucose uptake and lactate production by blocking the expression of GLUT4 in these cells. In addition, we found that SOX5 predominantly rescued the inhibitory effect of apatinib on GLUT4 expression by activating its promoter. Finally, we found that apatinib regulated the expression of SOX5 by suppressing the VEGFR2/AKT1/GSK3β signaling pathway.

Conclusions

From our results, we conclude that apatinib suppresses the in vitro and in vivo viability and proliferation of ovarian cancer cells, as well as glycolysis by inhibiting the VEGFR2/AKT1/GSK3β/SOX5/GLUT4 signaling pathway. Apatinib may serve as a promising drug for the treatment of ovarian cancer.

中文翻译：

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阿帕替尼通过抑制卵巢癌细胞中的VEGFR2 / AKT1 / SOX5 / GLUT4信号通路来抑制糖酵解。

背景

Apatinib是一种靶向血管内皮生长因子受体2（VEGFR2）的酪氨酸激酶抑制剂，已在各种恶性肿瘤中显示出令人鼓舞的治疗效果。然而，迄今为止，阿帕替尼在卵巢癌中的作用仍然未知。在这里，我们试图阐明阿帕替尼在卵巢癌细胞的体外和体内生存力和增殖以及这些细胞的葡萄糖代谢中的作用。

方法

分别使用Cell Counting Kit-8（CCK-8）和集落形成测定法评估了apatinib对卵巢癌细胞存活力和增殖的影响。使用蛋白质印迹法评估VEGFR2 / AKT1 / SOX5 / GLUT4途径蛋白的表达，并使用葡萄糖摄取和乳酸产生分析法检测卵巢癌细胞中的糖酵解。使用表达载体和基于shRNA的方法分别在卵巢癌细胞中外源性过表达SOX5和使其沉默。使用RNA-seq和基于基因芯片的方法进行RNA表达分析。GLUT4启动子活性使用双重荧光素酶报告基因分析进行评估。使用免疫组织化学（IHC）评估p-VEGFR2（Tyr1175），p-AKT1（Ser473），p-GSK3β（Ser9），SOX5和GLUT4在异种移植组织中的表达。

结果

我们发现，阿帕替尼以剂量依赖性和时间依赖性方式抑制Hey和OVCA433卵巢癌细胞的体外和体内生存力和增殖。我们还发现，阿帕替尼可通过阻断GLUT4在这些细胞中的表达来有效抑制葡萄糖的摄取和乳酸的产生。此外，我们发现SOX5主要通过激活启动子来挽救阿帕替尼对GLUT4表达的抑制作用。最后，我们发现阿帕替尼通过抑制VEGFR2 / AKT1 /GSK3β信号通路来调节SOX5的表达。

结论

根据我们的结果，我们得出结论，阿帕替尼通过抑制VEGFR2 / AKT1 /GSK3β/ SOX5 / GLUT4信号通路来抑制卵巢癌细胞的体外和体内生存力和增殖以及糖酵解。阿帕替尼可能是治疗卵巢癌的有前途的药物。