6-姜烯酚介导Hedgehog/Gli1通路对三阴性乳腺癌细胞侵袭及迁移作用机制

Effect of 6-Shogaol on invasion and migration of triple negative breast cancer cells via Hedgehog/Gli1 signaling pathway

目的研究Hedgehog(Hh)通路转录因子Gli1表达及6-姜烯酚介导Hh/Gli1通路对三阴性乳腺癌MDA-MB-231细胞增殖、侵袭及迁移能力的影响及分子机制。方法应用慢病毒建立过表达Gli1基因MDA-MB-231稳转细胞株;qRT-PCR及Western blot实验验证Gli1过表达效率;CCK-8和EdU实验检测Gli1表达及6-姜烯酚对细胞活力的影响;细胞划痕实验及Transwell实验检测细胞迁移和侵袭能力;Western blot实验检测Hh通路相关基因蛋白表达水平。结果成功建立MDA-MB-231-Gli1过表达稳转细胞株,当Gli1基因过表达时,细胞侵袭迁移能力明显提高,MDA-MB-231细胞中Hh通路基因Gli1,EMT标志基因Vimentin及Hippo通路基因YAP、TEAD4表达升高;应用Hh/Gli通路抑制剂Gant61抑制Gli1表达时,增殖、侵袭、迁移能力降低。应用6-姜烯酚处理细胞其增殖及侵袭迁移能力减弱,且Hh通路基因Gli1、Vimentin、YAP、TEAD4蛋白表达降低。结论Gli1基因能促进MDA-MB-231细胞侵袭迁移能力;6-姜烯酚能够通过Hedgehog信号通路抑制乳腺癌细胞增殖、侵袭迁移能力,转录因子Gli1可能是其作用靶点之一。

Aim To explore the effects of the expression of the transcription factor Gli1 of Hedgehog(Hh)signaling pathway and the 6-Shogaol mediated Hedgehog/Gli1 pathway on the proliferation,invasion and migration in MDA-MB-231 cells of triple negative breast cancer.Methods MDA-MB-231 cells were transfected by lentiviral vectors to stably overexpressed Gli1 gene.The overexpression efficiency of Gli1 was verified by qRT-PCR and Western blot.CCK-8 and EdU assays were used to detect the effect of Gli1 expression and 6-Shogaol on cell viability.Cell scratch assay and Transwell assay were used to detect the ability of migration and invasion.Western blot was used to detect the proteins expression of Hedgehog signaling pathway and other related genes.Results MDA-MB-231-Gli1 overexpression cell line was successfully established.When Gli1 gene was overexpressed,the invasion and migration ability of cells was significantly improved,and the expression of Hh signaling pathway gene Gli1,EMT marker gene Vimentin,Hippo signaling pathway genes YAP and TEAD4 increased.When the expression of Gli1 was inhibited by the Hh/Gli pathway inhibitor Gant61,the proliferation,invasion and migration abilities were suppressed.When the cells were treated with 6-Shogaol,the abilities of proliferation,invasion and migration were inhibited as well as the proteins expression of Gli1,Vimentin,YAP and TEAD4 decreased.Conclusions Gli1 gene can promote the invasion and migration of MDA-MB-231 cells.6-Shogaol can inhibit proliferation,invasion and migration of breast cancer cells through Hedgehog signaling pathway,suggesting that transcription factor Gli1 may be one of the targets of 6-Shogaol.