淫羊藿苷对表皮生长因子诱导的乳腺癌细胞MCF-7上皮间质化调控过程的影响

Effects of icariin on EGF-induced breast cancer cell MCF-7 epithelial mesenchymal regulation process and its mechanism

目的:探讨淫羊藿苷(ICA)对乳腺癌细胞MCF-7的作用,以及ICA对表皮生长因子(EGF)诱导的MCF-7细胞上皮间质转化(EMT)的作用及机制。方法:CCK-8法检测ICA对乳腺癌细胞MCF-7存活能力的影响并确定后续试验的干预浓度;设立空白对照组、EGF刺激组、EGF+ICA组;划痕实验、Transwell迁移及侵袭实验检测EGF及ICA+EGF对MCF-7细胞迁移及侵袭能力的影响;Western Blot检测AKT、p-AKT、N-cadherin、E-cadherin、Vimentin蛋白表达水平。结果:ICA对乳腺癌细胞MCF-7有杀伤作用,且呈浓度依赖性;各组MCF-7细胞24h愈合率由高到低依次为:EGF刺激组>EGF+ICA组>空白对照组(P<0.01);各组穿过膜的细胞数由高到低依次为:EGF刺激组>EGF+ICA组>空白对照组。与空白对照组比较,EGF刺激组p-AKT、N-cadherin、Vimentin蛋白表达量显著增加(P<0.01),E-cadherin蛋白表达量显著减少(P<0.01)。与EGF刺激组比较,EGF+ICA组p-AKT、N-cadherin、Vimentin蛋白表达量显著减少(P<0.01),E-cadherin蛋白表达量显著增加(P<0.01)。结论:ICA能够抑制乳腺癌细胞MCF-7的增殖。同时ICA还可以抑制EGF所诱导的EMT过程,可能是通过下调PI3K/AKT信号通路实现的。

Objective:To explore the effects of icariin(ICA)on breast cancer cells MCF-7,and the effect and mechanism of ICA on epithelial-mesenchymal transition(EMT)of MCF-7 cells induced by epidermal growth factor(EGF).Methods:CCK-8 method was used to detect the effects of ICA on the viability of breast cancer cells MCF-7 and determine the intervention concentration in subsequent experiments;set up blank control group,EGF stimulation group,EGF+ICA group;scratch test,Transwell migration and invasion test to detect the influence of EGF and ICA+EGF on the migration and invasion ability of MCF-7 cells;Western Blot detected the protein expression levels of AKT,p-AKT,N-cadherin,E-cadherin and Vimentin.Results:ICA had a killing effect on breast cancer cells MCF-7 and was concentration-dependent.The 24 h healing rate of MCF-7 cells in each group was as follows:EGF stimulation group>EGF+ICA group>blank control group(P<0.01);the number of cells passing through the membrane in each group was as follows:EGF stimulation group>EGF+ICA group>blank control group.Compared with the blank control group,the expression of p-AKT,N-cadherin and Vimentin in the EGF stimulation group increased significantly(P<0.01),and the expression of E-cadherin protein decreased significantly(P<0.01).Compared with the EGF stimulation group,the expression of p-AKT,N-cadherin and Vimentin in the EGF+ICA group was significantly reduced(P<0.01),and the expression of E-cadherin protein was significantly increased(P<0.01).Conclusion:ICA can inhibit the proliferation of breast cancer cell MCF-7.At the same time,ICA can also inhibit the EMT process induced by EGF,possibly by down-regulating the PI3 K/AKT signaling pathway.