摘要
目的检测叉头框架蛋白A2(FOXA2)在卵巢癌细胞中的表达情况,探讨其对上皮间质转化(EMT)的抑制作用及机制。方法培养卵巢癌上皮细胞系OVCAR3,构建并转染FOXA2过表达载体。采用CCK-8法、Transwell实验分别检测FOXA2过表达对OVCAR3细胞增殖、侵袭能力的影响;采用Western blot法及免疫荧光实验检测FOXA2过表达后OVCAR3细胞EMT发生相关蛋白E-钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)以及Wnt/β-catenin信号通路相关蛋白β-catenin的改变。结果FOXA2过表达明显抑制了OVCAR3细胞的增殖、侵袭能力。OVCAR3细胞FOXA2过表达后显著上调了E-cadherin蛋白的表达,下调了Vimentin蛋白的表达,即EMT过程受到了抑制。免疫荧光结果显示FOXA2过表达后OVCAR3细胞中β-catenin蛋白在细胞质和细胞核中出现明显的异位聚集。结论FOXA2过表达可能通过Wnt/β-catenin信号通路影响了EMT过程,进而抑制了卵巢癌OVCAR3细胞的增殖和侵袭能力。
Objective To investigate the effect of FOXA2 on epithelial-mesenchymal transformation(EMT)in ovarian epithelial carcinoma(EOC)cells and its related mechanism.Methods The FOXA2 overexpression vector was constructed and transfected to ovarian cancer epithelial OVCAR3 cells.CCK-8 and Transwell assay were used to detect the proliferation and invasion ability of FOXA2-overexpressing OVCAR3 cells.Western blot and immunofluorescence assay were used to detect the expression of E-cadherin,Vimentin and Wnt/β-catenin signaling pathway proteinβ-catenin after FOXA2 overexpression.Results FOXA2 overexpression significantly up-regulated the expression of E-cadherin and down-regulated the expression of Vimentin,indicating that the EMT process was inhibited.Immunofluorescence results showed thatβ-catenin protein in OVCAR3 cells was heterotopic in cytoplasm and nucleus after FOXA2 overexpression.Conclusion FOXA2 overexpression may affect EMT process through Wnt/β-catenin signaling pathway,and thereby inhibiting the proliferation and invasion of OVCAR3 cells.
作者
王凯
郭盼盼
管陈安
孙依娜
王俊强
余军辉
WANG Kai;GUO Panpan;GUAN Chenan;SUN Yina;WANG Junqiang;YU Junhui(Department of Obstetrics and Gynecology,Taizhou Hospital of Zhejiang Province affiliated to Wenzhou Medical University,Linhai 317000,China)
出处
《浙江医学》
CAS
2021年第20期2169-2174,I0002,共7页
Zhejiang Medical Journal
基金
浙江省公益技术应用研究计划项目(LGF19H160019)。
