淫羊藿苷通过Notch1/Jagged1信号通路抑制人前列腺癌细胞的增殖、迁移和侵袭

Inhibition of the proliferation,migration and invasion of prostate cancer cells by icariin via Notch1/Jagged1 pathway

目的:探讨淫羊藿主要有效成分淫羊藿苷(icariin,ICA)对人前列腺癌细胞(prostate cancer cells,PC-3细胞)的侵袭、迁移和增殖的影响及作用机制。方法:运用不同浓度的ICA对PC-3细胞进行24,36,48 h的预处理后,采用MTT法筛选合适的受试浓度;采用流式细胞术检测ICA对PC-3细胞周期的影响;采用划痕实验和Transwell实验分别检测ICA对PC-3细胞的迁移能力和侵袭能力的影响;采用RT-PCR和Western blot分别检测ICA对PC-3细胞Notch1、Jagged1、Hes1、Hey1 mRNA及蛋白表达的调控作用。结果:不同浓度的ICA处理PC-3细胞24,36,48 h后,对PC-3细胞的增殖有明显抑制作用,且呈浓度和时间依赖性;流式细胞术检测结果表明ICA主要将PC-3阻滞在G0/G1期;划痕实验和Transwell实验结果显示ICA对PC-3细胞的迁移和侵袭能力有明显抑制作用;1×10^(-4)mol·L^(-1)的ICA作用于PC-3细胞48 h后,对Notch1、Jagged1、Hes1、Hey1 mRNA表达有明显抑制作用(P<0.05),对Notch1、Jagged1蛋白及Hes1、Hey1蛋白表达均有抑制作用(P<0.01,P<0.05)。结论:淫羊藿主要有效成分ICA可以抑制PC-3细胞的迁移、侵袭和增殖,其机制可能与下调Notch1/Jagged1通路的相关因子有关。

OBJECTIVE To observe the effects and mechanism of icariin(ICA),the main active ingredient of Epimedium,on the invasion,migration and proliferation of human prostate cancer cells(PC-3 cell)in vitro.METHODS After pretreating PC-3 cells with different concentrations of ICA for 24,36,and 48 h,MTT method screened for appropriate subject concentrations;Flow cytometry to detect the effect of ICA on the PC-3 cell cycle;the migration abilities of PC-3 cells effected by ICA were observed by scratch assay;the invasion abilities of PC-3 cells effected by ICA were observed by Transwell assay.RT-PCR and Western blot respectively detected the regulation of ICA on Notch1,Jagged1,Hes1,Hey1 mRNA and protein expression in PC-3 cells.RESULTS After treating PC-3 cells at different concentrations of ICA for 24,36 and 48 h,ICA significantly inhibited the proliferation of PC-3 cells,and it was concentration and time dependent.The flow cytometry result showed that ICA mainly blocked PC-3 in the G0/G1 stage.The result of scratch assay experiment showed that ICA significantly inhibited the migration ability of PC-3 cells.The result of Transwell experiment also showed that ICA significantly inhibited the invasion ability of PC-3 cells.ICA concentration of 1×10^(-4) mol·L^(-1) acted on PC-3 cells for 48 h,and had a significant inhibitory effects on Notch1,Jagged1 and Hes1,Hey1 mRNA and the protein expression levels of Hes1 and Hey1(P<0.05);and had a very significant inhibitory effects on protein expression levels of Notch1 and Jagged1(P<0.01).CONCLUSION ICA,the main active ingredient of Epimedium,could inhibit the proliferation,migration and invasion of PC-3 cells,the mechanism may be related to the down-regulation of related factors of Notch1/Jagged1 pathway.