毛萼乙素通过Wnt/β-catenin通路抑制结肠癌细胞上皮间质转化

Inhibitory effects of eriocalyxin B on epithelial-mesenchymal transition in colon cancer cells through the Wnt/β-catenin pathway

目的 研究毛萼乙素对结肠癌细胞上皮间质转化(EMT)的影响及机制。方法 以结肠癌细胞HT29、HCT116为研究对象。采用划痕愈合实验和Transwell实验检测毛萼乙素对2种细胞迁移和侵袭能力的影响;采用Western blot法检测毛萼乙素(1.0μmol/L)、毛萼乙素+Wnt/β-联蛋白(β-catenin)信号通路激动剂Licl(1.0μmol/L毛萼乙素+10 mmol/L Licl)或毛萼乙素+Wnt/β-catenin信号通路抑制剂XAV939(1.0μmol/L毛萼乙素+10μmol/L XAV939)对2种细胞的EMT相关蛋白[E-钙黏蛋白(Ecadherin)、N-钙黏蛋白(N-cadherin)、锌指转录抑制因子Snail]和Wnt/β-catenin信号通路相关蛋白[β-catenin、原癌基因蛋白质cmyc、细胞周期蛋白D1(cyclin D1)、T细胞因子4(TCF4)]表达的影响。结果 毛萼乙素可显著抑制2种细胞的侵袭和迁移(P<0.01),并显著降低2种细胞中N-cadherin、Snail、β-catenin、c-myc、cyclin D1、TCF4表达水平,显著升高E-cadherin表达水平(P<0.05或P<0.01)。毛萼乙素与Licl联用后,2种细胞中β-catenin、c-myc、cyclin D1、TCF4及N-cadherin表达水平被逆转(P<0.05或P<0.01);与XAV939联用后,2种细胞中β-catenin、N-cadherin的表达进一步下调(P<0.05或P<0.01),E-cadherin的表达进一步上调(P<0.01),且细胞迁移率也显著降低(P<0.01)。结论 毛萼乙素可通过抑制Wnt/β-catenin通路激活抑制结肠癌细胞上皮间质转化。

OBJECTIVE To study the effects and mechanism of eriocalyxin B(EriB) on epithelial-mesenchymal transition(EMT)of colon cancer cells.METHODS The colon cancer cells HT29 and HCT116 as research objects.Scratch-healing assay and Transwell assay were used to detect the effects of EriB on migration and invasion of two kinds of cells.Western blot method was used to detect the effects of EriB(1.0 μmol/L),EriB+Wnt/β-catenin signaling pathway agonist Licl(1.0 μmol/L EriB+10 mmol/L Licl) or EriB+Wnt/β-catenin signaling pathway inhibitor XAV939(1.0 μmol/L EriB+10 μmol/L XAV939) on the expressions of EMT related proteins [E-cadherin,N-cadherin,Snail] and Wnt/β-catenin signaling pathway related proteins [β-catenin,c-myc,cyclin D1,TCF4].RESULTS EriB can significantly inhibit the invasion and migration of two kinds of cells(P<0.01).EriB significantly reduced the protein expressions of N-cadherin,Snail,β-catenin,c-myc,cyclin D1 and TCF4 while increases the protein expression of E-cadherin(P<0.05 or P<0.01).After EriB combined with Licl,the protein expressions of β-catenin,cmyc,cyclin D1 and TCF4 in the two kinds of cells were reversed(P<0.05 or P<0.01).After combined with XAV939,the protein expressions of β-catenin and N-cadherin were further down-regulated(P<0.05 or P<0.01);the protein expression of E-cadherin was further up-regulated(P<0.01),while migration rate was decreased significantly(P<0.01).CONCLUSIONS Eri B can inhibit the epithelial-mesenchymal transition of colon cancer cells by inhibiting the activation of Wnt/β-catenin pathway.