和厚朴酚对宫颈癌细胞血管生成拟态及上皮间质转化的影响

Effects of honokiol on vasculogenic mimicry and epithelial-mesenchymal transition of cervical cancer cells

目的探讨和厚朴酚调节表皮生长因子受体(epithelial growth factor receptor,EGFR)信号通路对宫颈癌细胞血管生成拟态(vasculogenic mimicry,VM)和上皮间质转化(epithelial-mesenchymal transition,EMT)的影响。方法将宫颈癌HeLa细胞分为对照组、和厚朴酚低、中、高剂量组、和厚朴酚+NSC228155组以及西妥昔单抗组;采用Transwell小室检测细胞侵袭;划痕试验检测细胞迁移;三维培养HeLa细胞并进行高碘酸希夫染色,观察VM形成情况;采用蛋白质印迹法和实时荧光定量聚合酶链反应检测细胞中的蛋白和基因表达。结果与对照组比较,和厚朴酚低、中、高剂量组HeLa细胞侵袭数目、划痕愈合率、环状结构数、磷酸化EGFR(phosphorylation EGFR,p-EGFR)/EGFR、基质金属蛋白酶(matrix metalloproteinase,MMP)2、MMP9、促红细胞生成素肝细胞受体(erythropoietin-producing hepatocyte receptor,Eph)A2、血管内皮生长因子(vascular endothelial growth factor,VEGF)、神经钙黏蛋白(neural cadherin,CDH2)以及波形蛋白表达均降低,而上皮钙黏蛋白(epithelial cadherin,CDH1)表达升高(P<0.01);与和厚朴酚高剂量组比较,和厚朴酚+NSC228155组HeLa细胞侵袭数目、划痕愈合率、环状结构数、p-EGFR/EGFR、MMP2、MMP9、EphA2、VEGF、CDH2以及波形蛋白表达均升高,而CDH1表达降低(P<0.01)。结论和厚朴酚可能通过抑制EGFR信号通路抑制宫颈癌细胞VM形成和EMT。

Objective To investigate the effects of honokiol on vasculogenic mimicry(VM)and epithelial-mesenchymal transition(EMT)of cervical cancer cells by regulating epidermal growth factor receptor(EGFR)signaling pathway.Methods HeLa cervical cancer cells were divided into control group,honokiol low-dose,medium-dose and high-dose groups,honokiol+NSC228155 group,and cetuximab group.Transwell chambers were used to detect cell invasion.Scratch assay was applied to detect cell migration.Periodic acid-Schiff was used to observe the formation of VM in HeLa cells.Western blot and quantitative real time fluorescent polymerase chain reaction were applied to detect the protein and gene expression in cells.Results Compared with the control group,the number of HeLa cell invasion,scratch healing rate,number of ring structures,the ratio of phosphorylated EGFR(p-EGFR)/EGFR,the expression of matrix metalloproteinase(MMP)2,MMP9,erythropoietin-producing hepatocyte receptor(Eph)A2,vascular endothelial growth factor(VEGF),neural cadherin(CDH2)and vimentin in the low,medium and high dose honokiol groups were decreased,and the expression of epithelial cadherin(CDH1)was increased(P<0.01).Compared with the high-dose honokiol group,the number of invasion,scratch healing rate,number of ring structures,the ratio of p-EGFR/EGFR,the expression of MMP2,MMP9,EphA2,VEGF,CDH2 and Vimentin in the honokiol+NSC228155 group were increased,and the expression of CDH1 was decreased(P<0.01).Conclusion Honokiol may inhibit VM formation and EMT of cervical cancer cells by inhibiting the EGFR signaling pathway.