EphA5 knockdown enhances the invasion and migration ability of esophageal squamous cell carcinoma via epithelial-mesenchymal transition through activating Wnt/β-catenin pathway
EphA5 knockdown enhances the invasion and migration ability of esophageal squamous cell carcinoma via epithelial-mesenchymal transition through activating Wnt/β-catenin pathway
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DOI：
10.1186/s12935-020-1101-x
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发表时间：
2020-01-13
影响因子：
5.8
通讯作者：
Zhou, Shao-Bing
中科院分区：
文献类型：
--
作者：
Zhang, Rui;Liu, Jing;Zhou, Shao-Bing
背景 促红细胞生成素生成肝细胞 (Eph) 受体 A5 (EphA5) 已被发现在一些恶性肿瘤中过度表达，并与疾病预后相关。然而，EphA5在食管鳞状细胞癌（ESCC）中的作用尚不清楚。方法在本研究中，我们测量了ESCC组织和细胞系（包括KYSE150和KYSE450细胞）中EphA5的表达。 siRNA转染用于干扰ESCC细胞系中EphA5的表达。进行细胞活力、集落形成、划痕和侵袭测定，以探索 EphA5 在 ESCC 细胞系中的作用。流式细胞术分析EphA5是否影响细胞凋亡和周期。还通过蛋白质印迹和免疫荧光测量了与上皮间质转化 (EMT) 相关的生物标志物和与 Wnt/β-连环蛋白信号传导相关的分子。结果新鲜ESCC组织和细胞系中EphA5蛋白和mRNA的表达量显着高于正常对照组和人正常食管上皮细胞（HEEC）。细胞活力测定和集落形成测定表明，EphA5 敲低增强了 KYSE150 和 KYSE450 细胞的体外增殖。 EphA5敲低后ESCC细胞的侵袭和迁移加速。在转染针对 EphA5 的 siRNA 的 ESCC 细胞中，EMT 生物标志物的表达发生了改变。此外，EphA5 下调增强了 β-catenin 和 p-GSK-3 beta(Ser9) 的蛋白水平，它们在 Wnt/β-catenin 通路中发挥着关键作用。结论 EphA5敲低可促进食管鳞癌的增殖，并通过激活Wnt/β-catenin通路，通过上皮间质转化增强侵袭和迁移能力。
Background The erythropoietin-producing hepatocellular (Eph) receptor A5 (EphA5) has been found to be overexpressed in some malignant tumors and is associated with disease prognosis. However, the role of EphA5 in esophageal squamous cell carcinoma (ESCC) is not clear. Methods In the present study, we measured the expression of EphA5 in ESCC tissues and cell lines including KYSE150 and KYSE450 cells. siRNA transfection was used to interfere with EphA5 expression in ESCC cell lines. Cell viability, colony formation, scratch and invasion assays were performed to explore the roles of EphA5 in ESCC cell lines. Flow cytometry analysis was performed to investigate whether EphA5 could affect the cell apoptosis and cycle. The biomarkers related to epithelial-mesenchymal transition (EMT) and molecules associated with Wnt/beta-catenin signaling were also measured by western blot and immunofluorescence. Results The protein and mRNA expression of EphA5 were significantly higher in fresh ESCC tissues and cell lines compared with normal control groups and human normal esophageal epithelial cells (HEEC). The cell viability assay and colony formation assay revealed that EphA5 knockdown enhanced the proliferation of KYSE150 and KYSE450 cells in vitro. The invasion and migration of ESCC cells were accelerated after EphA5 knockdown. The expression of EMT biomarkers was altered in ESCC cells transfected with siRNA targeting EphA5. Moreover, EphA5 downregulation enhanced the protein levels of beta-catenin and p-GSK-3 beta(Ser9), which play a key role in the Wnt/beta-catenin pathway. Conclusions EphA5 knockdown promotes the proliferation of esophageal squamous cell carcinoma,enhances invasion and migration ability via epithelial-mesenchymal transition through activating Wnt/beta-catenin pathway.