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目的:探讨金荞麦提取物对WM239细胞穿越内皮细胞过程中N-钙黏附分子的影响。方法:采用苏木素方法测定金荞麦提取物对HUVEC和WM239细胞增殖的影响,筛选合适的金荞麦提取物浓度;利用Transwell迁移实验,测定金荞麦提取物对WM239细胞迁移能力的影响;流式细胞术(FCM)测定金荞麦提取物对HUVEC和WM239细胞共培养体系N-cadherin表达的影响;免疫共沉淀实验测定金荞麦提取物对N-cadherin胞内段磷酸化及其与β-catenin结合的影响;Western blot检测金荞麦提取物对Src活化的影响。结果:金荞麦提取物浓度在25～100 mg/L之间对WM239和HUVEC的增殖无明显抑制作用。金荞麦提取物在此药物浓度范围内可对WM239细胞的迁移能力有较强的抑制作用。金荞麦提取物对共培养细胞N-cadherin表达量几乎没有影响,但可导致p-Src蛋白的表达减少,N-cadherin胞内段的磷酸化减低,引起N-cadherin与β-catenin的解离减少。结论:金荞麦提取物可抑制WM239的增殖和迁移能力,减少Src蛋白的活化,降低N-cadherin胞内段磷酸化水平及N-cadherin与β-catenin的解离。
Objective: To investigate the effect of F buckle on the expression of N-cadherin in the process of WM239 cells crossing the endothelial cells. Methods: Hematoxylin was used to determine the effect of F buckle extract on the proliferation of HUVEC and WM239 cells, and the appropriate concentration of F buckle extract was screened. The effect of F buckle extract on the migration of WM239 cells was determined by Transwell migration assay. Flow cytometry (FCM) was used to determine the effect of FBS on the expression of N-cadherin in co-culture system of HUVEC and WM239 cells; the effect of FBS on phosphorylation of N-cadherin and its interaction with β-catenin Western blot was used to detect the effect of F buckle extract on Src activation. Results: F buckle extract concentration between 25 ~ 100 mg / L had no obvious inhibitory effect on the proliferation of WM239 and HUVEC. The buckwheat extract can inhibit the migration of WM239 cells in this drug concentration range. However, the expression of p-Src decreased and the phosphorylation of intracellular N-cadherin decreased, which led to the dissociation of N-cadherin and β-catenin cut back. Conclusion: Buckwheat extract can inhibit the proliferation and migration of WM239, reduce the activation of Src protein, decrease the intracellular phosphorylation of N-cadherin and the dissociation of N-cadherin and β-catenin.