异泽兰黄素调节Notch 1/Hes信号通路对宫颈癌细胞迁移和侵袭的影响

The effects of Eupatilin on the migration and invasion of cervical cancer cells by regulating Notch 1/Hes signaling pathway

目的探讨异泽兰黄素对人宫颈癌(cervical cancer,CC)细胞迁移和侵袭的影响及对Notch信号通路的调节作用。方法培养人CC肠转移细胞系Caski细胞,将其分为空白对照(BC)组、阴性对照(NC)组、异泽兰黄素(Eupatilin)组、Notch 1过表达(Notch 1)组及异泽兰黄素+Notch 1过表达(Eupatilin+Notch 1)组。MTT法检测细胞活力;划痕愈合实验与Transwell实验检测细胞迁移和侵袭情况;蛋白质印迹法检测细胞中上皮间质转化(epithelial–mesenchymal transition,EMT)和Notch信号通路相关蛋白水平。结果与NC组相比,Eupatilin组细胞活力、迁移、侵袭能力以及细胞中N-cadherin、Snail、Notch 1、Hes-1及Hes-5蛋白水平均降低,细胞中E-cadherin蛋白水平增高(P<0.05);Notch 1组以上各指标变化与Eupatilin组呈相反的效果(P<0.05)。Notch 1过表达逆转异泽兰黄素对Caski细胞活力、迁移、侵袭以及Notch信号通路的影响(P<0.05)。结论异泽兰黄素通过Notch 1/Hes信号通路抑制人CC细胞迁移和侵袭,该研究为人类CC的临床治疗提供一些新的线索。

Objective To investigate the effects of Eupatilin on the migration and invasion of human cervical cancer(CC)cells and its regulation on Notch signaling pathway.Methods The Caski cells,a human CC intestinal metastasis cell line,were cultured and divided into blank control(BC)group,negative control(NC)group,Eupatilin group,Notch 1 overexpression(Notch 1)group,Eupatilin and Notch 1 overexpression(Eupatilin+Notch 1)group.MTT method was performed to measure the viability of cells.Scratch healing test and Transwell test were performed to measure the migration and invasion of cells.Western blot was performed to measure the levels of epithelial-mesenchymal transition(EMT)and Notch signaling pathway-related proteins in cells.Results Compare with the NC group,the cell viability,migration and invasion abilities,N-cadherin,Snail,Notch 1,Hes-1 and Hes-5 protein levels in Eupatilin group were decreased,and the E-cadherin protein level was increased(P<0.05).The changes of the above indexes in Notch 1 group were opposite to those in Eupatilin group(P<0.05).The overexpression of Notch 1 reversed the effects of Eupatilin on the viability,migration and invasion of Caski cells and the Notch signaling pathway(P<0.05).Conclusion Eupatilin inhibits the migration and invasion of human CC cells through Notch 1/Hes signaling pathway,which may provide some new clues for the clinical treatment of human CC.