XAV939对结肠癌耐药细胞迁移能力的影响

Effects of XAV939 on migration ability of drug-resistant colon cancer cells

目的探究Wnt通路抑制剂XAV939对结肠癌耐药细胞迁移能力的影响。方法 XAV939处理HCT-8/5-Fu细胞,MTT检测HCT-8/WT、HCT-8/5-Fu和HCT-8/5-Fu+XAV939细胞对5-氟尿嘧啶(5-fluorouracil,5-Fu)的敏感性;Transwell和划痕实验检测细胞迁移能力;Western blot检测β-catenin、c-Myc和cyclin D1蛋白的表达水平。结果 MTT、Transwell和划痕实验结果显示,HCT-8/5-Fu细胞对5-Fu的敏感性低于HCT-8/WT细胞,迁移能力高于HCT-8/WT细胞;Western blot结果显示,与HCT-8/WT细胞相比,HCT-8/5-Fu细胞核中β-catenin明显高表达,且c-Myc和cyclin D1也呈现高表达;XAV939处理HCT-8/5-Fu细胞后,其对5-Fu的耐受性和迁移能力均降低,细胞核中β-catenin表达下降,并且c-Myc和cyclin D1的表达也明显减少。结论 Wnt/β-catenin通路可能参与了HCT-8/5-Fu细胞的耐药,其小分子抑制剂XAV939通过影响HCT-8/5-Fu细胞对5-Fu的耐受性,进而影响细胞的迁移能力。

Aim To explore the effect of Wnt pathway inhibitor XAV939 on migration ability of drug-resistant colon cancer cells. Methods HCT-8/5-Fu cells were treated with XAV939. MTT was used to detect the sensitivity of HCT-8/WT,HCT-8/5-Fu and HCT-8/5-Fu＋ XAV939 cells to 5-fluorouracil（5-Fu）. Transwell and wound healing assay were used to detect the ability of cell migration. The protein levels of β-catenin,cMyc and cyclin D1 were validated by Western blot.Results The results of MTT assay,Transwell and wound healing assay showed that the sensitivity of HCT-8/5-Fu cells to 5-Fu was lower than that of HCT-8/WT cells,and the ability of migration was stronger than that of HCT-8/WT cells. Western blot showed β-catenin expressed higher in the nucleus of HCT-8/5-Fu cells than in HCT-8/WT cells. Moreover,we found c-Myc and cyclin D1 were highly expressed in HCT-8/5-Fu cells than in HCT-8/WT cells. Importantly,when treated HCT-8/5-Fu cells with XAV939,the sensitivity to 5-Fu,the ability of migration,the expression of β-catenin,c-Myc and cyclin D1 significantly decreased.Conclusions Wnt/β-catenin pathway may be involved in the drug resistance of colon cancer cells,and its small molecular inhibitor,XAV939,may reduce cell migration through affecting the resistance of HCT-8/5-Fu cells to 5-Fu.