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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。
AIM: To investigate the role of Hedgehog (HH) signaling pathway effector protein GLI-1 in enhancing epidermal growth factor (EGF) -mediated invasion of human prostate cancer AR-CaPE cells in vitro. Methods: Human prostate cancer ARCaP cell line was used as a model to identify the expression of EGFR and GLI-1 in ARCaPE by immunofluorescence staining. The effect of 100 ng / ml EGF on ARCaPE was observed in vitro and in vitro (100 ng / ml) and GLI-1 antagonist GANT61 (10μmol / L) alone or in combination with GLUT-1 (10μmol / L) Effect of Combined Action on In Vitro Invasion Ability of ARCaPE Cells. Results: ARCaPE cells expressed EGFR and GLI-1 protein simultaneously. EGF induced epithelial-like ARCaPE cells to transform into stromal-like AR-CaPM, enhanced the invasiveness of ARCaPE cells in vitro and up-regulated the expression of p-ERK and GLI- 1 protein (P <0.05). GANT61 significantly inhibited the invasiveness of ARCaPE cells in vitro and attenuated the enhanced effect of EGF on cell invasiveness (P <0.05). CONCLUSION: There is a certain interaction between HH signaling pathway and EGF / ERK signaling pathway. GLI-1 may play an important role in EGF-mediated enhancement of invasiveness of human prostate cancer ARCaPE cells in vitro.