摘要
目的:探讨Hedgehog(HH)信号通路效应蛋白GLI-1在表皮生长因子(EGF)介导的人前列腺癌AR-CaPE细胞系体外侵袭活性增强中的作用。方法:以人前列腺癌ARCaP细胞系作为研究模型,免疫荧光技术鉴定ARCaPE内EGF受体(EGFR)和GLI-1的表达情况;100 ng/ml EGF体外作用于ARCaPE后,观察细胞的形态及体外侵袭能力的变化,采用Western印迹检测细胞内ERK信号通路成分和GLI-1蛋白的表达变化情况;Transwell侵袭实验检测EGF(100 ng/ml)与GLI-1拮抗剂GANT61(10μmol/L)单独或联合作用对ARCaPE细胞体外侵袭能力的影响。结果:ARCaPE细胞同时表达EGFR与GLI-1蛋白;EGF诱导上皮样外观的ARCaPE细胞向间质样外观的AR-CaPM转化,增强ARCaPE细胞的体外侵袭能力并显著上调细胞内p-ERK和GLI-1蛋白的表达水平(P<0.05);GANT61明显抑制ARCaPE细胞的体外侵袭能力且减弱EGF对细胞侵袭能力的增强效应(P<0.05)。结论:HH信号通路和EGF/ERK信号通路之间存在一定的相互作用,GLI-1可能在EGF介导的人前列腺癌ARCaPE细胞体外侵袭活性增强过程中发挥着重要作用。
Objective： To investigate the role of the hedgehog（HH） signaling pathway transcription factor glioma-associated oncogene hoinolog 1（GLI-1） in EGF-regulated enhancement of the invasiveness of the prostate cancer ARCaPE cell line in vitro.Methods： The expressions of EGFR and GLI-1 in prostate cancer ARCaPE cells were analyzed by immunofluorescence staining.ARCaPE cells were treated with EGF at 100 ng/ml,followed by detection of the changes in cell morphology and invasiveness,as well as in the expressions of p-ERK,ERK and GLI-1.Migration transwell assay was used to determine the effects of 100 ng/ml EGF and GLI-1 antagonist GANT61 on the invasiveness of the ARCaPE cells.Results： Both EGFR and GLI-1 were expressed in the ARCaPE cells.EGF induced morphological transition of epithelial-like ARCaPE cells to mesenchymal-like cells,increased their in vitro invasiveness,and significantly upregulated the expressions of p-ERK and GLI-1 in the ARCaPE cells（P〈0.05）.GANT61 significantly inhibited the in vitro invasiveness of the ARCaPE cells and reduced the enhancing effect of EGF on their invasiveness（P〈0.05）.Conclusion： The results from ARCaPE cells shed light on the cross-talk of the HH pathway with the EGF/ERK signaling pathway.GLI-1 might be responsible for EGF-regulated enhancement of the invasiveness of ARCaPE cells in vitro.
出处
《中华男科学杂志》
CAS
CSCD
2012年第1期16-22,共7页
National Journal of Andrology
基金
国家自然科学基金(30901501)~~
