白藜芦醇对人结肠癌细胞上皮间质转化的抑制作用

目的观察白藜芦醇(Resveratrol,Res)对人结肠癌细胞HCT-8上皮间-质转化(EMT)的影响,初步探讨其可能存在的机制。方法取对数生长期的人结肠癌HCT-8细胞,采用不同浓度的白藜芦醇(0、40、80、120μmol/L)对细胞分别进行干预24、48、72 h;MTT法检测不同时间Res对HCT-8细胞增殖活性的影响;细胞黏附试验检测Res对HCT-8细胞黏附能力的影响;Transwell小室试验检测Res对HCT-8细胞迁移和侵袭能力的影响;进一步采用蛋白免疫印迹法和qRT-PCR分别检测Res对HCT-8细胞中蛋白激酶B(AKT)、磷酸化蛋白激酶B(P-AKT)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和mRNA表达的影响。结果 Res(浓度为40、80、120μmol/L)在干预后24、48、72 h均可以明显抑制HCT-8细胞的增殖活性(P<0.05)。与空白对照组相比,细胞的同种黏附能力增强,异种黏附能力降低,差异有统计学意义(P<0.05)。与空白对照组相比,Res 120μmol/L对细胞迁移和侵袭能力均具有明显的抑制作用(P<0.05),Res 120μmol/L作用HCT-8细胞48 h后,HCT-8细胞中上皮间质标志性蛋白E-钙黏蛋白(E-cadherin)和mRNA的相对表达量增加(P<0.05),而AKT蛋白的表达无明显变化(P>0.05),P-AKT蛋白的表达明显减少(P<0.05);N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和mRNA的相对表达量显著减少,差异有统计学意义(P<0.05)。结论白藜芦醇通过调控细胞EMT抑制人结肠癌HCT-8细胞的迁移和侵袭能力,其作用机制可能受PI3K/AKT信号通路的调控。

Inhibitory effect of resveratrol on epithelial-mesenchymal transition in human colon cancer cells

Objective To observe the effect of resveratrol(Res)on the epithelial-mesenchymal transition of human colon cancer cell line HCT-8,and to explore its possible mechanism.Methods Human colon cancer HCT-8 cells in logarithmic growth phase were treated with different concentrations of resveratrol(0,40,80,120μmol/L)for 24 h,48 h and 72 h respectively.MTT assay was used to detect the effect of Res on the proliferation of HCT-8 cells at different time points.The effect of Res on the adhesion of HCT-8 cells was detected by cell adhesion assay.The effect of Res on the migration and invasion of HCT-8 cells was detected by Transwell chamber assay.The expression of AKT,P-AKT,E-cadherin,N-cadherin and Vimentin protein and mRNA in HCT-8 cells was detected by Western blot and qRT-PCR,respectively.Results Res(at concentration of 40,80,120μmol/L)significantly inhibited the proliferation of HCT-8 cells at 24 h,48 h and 72 h after intervention(P<0.05).Compared with blank control group,the ability of homologous cell adhesion increased and the ability of heterologous cell adhesion decreased(P<0.05).Compared with blank control group,Res 120μmol/L had significant inhibitory effect on HCT-8 migration and invasion ability(P<0.05).The relative expression of epithelial mesenchymal marker protein E-cadherin and mRNA in HCT-8 cells increased after Res 120μmol/L treated HCT-8 for 48 h(P<0.05),while the expression of AKT protein did not change significantly(P>0.05),and the expression of P-AKT protein was significantly decreased(P>0.05).The relative expression levels of N-cadherin,Vimentin protein and mRNA were significantly reduced(P<0.05).Conclusion Resveratrol inhibits the migration and invasion of human colon cancer HCT-8 cells by regulating cell epithelial-mesenchymal transition(EMT),and its mechanism may be regulated by PI3 K/AKT signaling pathway.