二氢杨梅素对人胃癌MKN45细胞迁移和侵袭的影响及其机制

Effects of dihydromyricetin on the migration and invasion of human gastric cancer MKN45 cells and its mechanism

目的:探讨二氢杨梅素(DHM)对人胃癌MKN45细胞迁移和侵袭的作用及其分子机制。方法:培养人低分化胃癌MKN45细胞,用不同浓度的DHM(0,10,20,30,40,50μmol/L)分别处理细胞24及48 h,每组实验重复3次,采用CCK8实验检测癌细胞增殖活力;划痕实验检测细胞迁移能力;Transwell小室检测细胞侵袭能力;免疫印迹分析细胞迁移和侵袭相关蛋白表达情况。结果:不同浓度DHM干预可降低MKN45细胞活力。20,30及40μmol/L的DHM处理48 h可明显抑制细胞的迁移能力(P<0.01)和侵袭能力(P<0.05及0.01)。20及30μmol/L的DHM处理48 h可增加E-cadherin蛋白表达(P<0.01)、降低Vimentin表达(P<0.01),从而逆转EMT过程;10,20及30μmol/L的DHM处理48 h可明显降低pJNK的活性表达水平(P<0.05及0.01),及MMP-2蛋白表达(P<0.01);JNK通路特异性抑制剂SP600125预处理可明显促进DHM对癌细胞侵袭能力的抑制作用(P<0.01)及降低MMP-2表达(P<0.01)。结论:DHM具有抑制人胃癌MKN45细胞的迁移及侵袭的作用,其机制可能与通过JNK通路下调MMP-2蛋白表达水平、逆转上皮间质转化有关。

Objective:To investigate the effects of dihydromyricetin(DHM)on the migration and invasion of human gastric cancer MKN45 cells and its mechanism and provide experimental basis for the prevention and treatment of gastric cancer with Traditional Chinese Medicine(TCM).Methods:MKN45 cells were pre-treated with DHM(0,10,20,30,40,50μmol/L)for 24 and 48 hours respectively.Cell viability treated with different concentrations of DHM was detected by Cell Counting kit(CCK-8)assay,cell migration was measured by wound healing assay,and cell invasion was tested by Transwell assay.Cells were pre-treated with DHM or co-treated with c-Jun N-terminal kinase(JNK)pathway inhibitor SP600125,then,the levels of migration-and invasion-related proteins were tested by Western blot.Results:DHM concentration-dependently inhibited cell migration and invasion and downregulated matrix metalloprotein-2(MMP-2)and phosphorylated JNK(pJNK)expression in MKN45 cells,followed by upregulation of E-cadherin and downregulation of Vimentin.Co-treatment with DHM and JNK inhibitor SP600125 further suppressed MMP-2 expression and cell invasion in MKN45 cells,suggesting that DHM inhibited MKN45 cells metastasis through JNK/MMP-2 pathway.Conclusion:DHM can inhibit cell migration and invasion in human gastric cancer MKN45 cells through downregulating MMP-2 expression via JNK signaling pathway and reverse epithelial-mesenchymal transition(EMT),implying that DHM could have the potential to serve as an anti-metastatic agent for treating gastric cancer.