安罗替尼靶向VEGFR2增强食管癌ECA-109细胞对紫杉醇的敏感性

Anlotinib Enhances the Sensitivity of Esophageal Cancer ECA-109 Cells to Paclitaxel by Targeting VEGFR2

目的探讨安罗替尼(Anlotinib)通过靶向血管内皮生长因子受体2(VEGFR2)增强紫杉醇对食管癌ECA-109细胞的增殖、侵袭和迁移、凋亡的影响以及可能的作用机制。方法通过免疫组化法检测VEGFR2在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中的表达及与临床病理特征的关系。CCK8法、流式细胞术、Transwell实验和细胞凋亡实验检测Anlotinib、Anlotinib联合紫杉醇对ECA-109细胞增殖、侵袭及迁移、凋亡的影响;蛋白质免疫印迹(Western blotting)检测磷酸化的哺乳动物雷帕霉素靶蛋白(p-mTOR)、磷酸化的蛋白激酶B(p-AKT)、磷酸化的血管内皮生长因子受体2(p-VEGFR2)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2-Associated X的蛋白质(Bax)、活性半胱氨酸天冬氨酸蛋白酶-3(active Caspase-3)的表达。结果VEGFR2在ESCC患者临床标本中呈高表达并且与TNM分期有关(P<0.05),Anlotinib联合紫杉醇较Anlotinib单药抑制ECA-109细胞的增殖、侵袭和迁移,促进了细胞凋亡(P<0.05);Western blotting检测结果显示,p-mTOR、p-AKT、P-VEGFR2、Bcl-2蛋白表达水平降低,Bax、Active Caspase-3蛋白表达水平升高。结论VEGFR2在ESCC患者临床标本中高表达且与TNM分期有关,是预后不良的指标。Anlotinib联合紫杉醇抑制ECA-109细胞的增殖、侵袭和迁移,促进了细胞凋亡。Anlotinib联合紫杉醇可能通过VEGFR2/AKT/mTOR信号通路发挥抑癌作用,为食管癌治疗提供理论依据。

Objective To investigate the effect of Anlotinib on the proliferation,invasion,migration and apoptosis of esophageal cancer ECA-109 cells by targeting vascular endothelial growth factor receptor 2(VEGFR2)and its possible mechanism.Methods The expression of VEGFR2 in esophageal squamous cell carcinoma and its relationship with clinicopathological features were detected by immunohistochemistry.The effects of Anlotinib,Anlotinib combined with paclitaxel on the proliferation,invasion,migration and apoptosis of ECA-109 cells were detected by CCK8 method,flow cytometry,Transwell assay and apoptosis assay.Western blotting was used to detect the expressions of phosphorylated mammalian cells Target of Rapamycin(p-mTOR),Phosphorylated Protein Kinase B(p-AKT),Phosphorylated Vascular Endothelial Growth Factor Receptor 2(p-VEGFR2),B Lymphoma-2(Bcl-2),Bcl-2-Associated X protein(Bax)and Active Caspase-3.Results VEGFR2 was highly expressed in patients with esophageal squamous cell carcinoma and was related to TNM staging(P<0.05).Compared with Anlotinib alone,Anlotinib combined with paclitaxel significantly inhibited the proliferation,invasion and migration of ECA-109 cells and promoted cell apoptosis(P<0.05).Western blotting results showed that p-mTOR,p-AKT,P-VEGFR2,Bcl-2 decreased and Bax,Active Caspase-3 increased.Conclusion VEGFR2 is an indicator of poor prognosis,which is highly expressed in patients with esophageal squamous cell carcinoma and associated with TNM staging.Anlotinib combined with paclitaxel can inhibit the proliferation,invasion and migration of ECA-109 cells,and promote cell apoptosis.Anlotinib combined with paclitaxel can provide a theoretical basis for the treatment of esophageal cancer,which may exert a tumor suppressor effect through the VEGFR2/AKT/mTOR signaling pathway.