XAV939对胃癌细胞株MKN-45的增殖及Wnt/β-连环蛋白信号通路的抑制作用

Inhibitory effects of XAV939 on gastric carcinoma MKN-45 cells proliferation and Wnt/β-catenin signaling pathway in vitro

目的 观察XAV939对胃癌细胞株MKN-45增殖及Wnt/β-连环蛋白（Wnt/β-catenin）信号通路的作用.方法 检测XAV939对胃癌细胞株MKN-45细胞的增殖抑制作用,逆转录-聚合酶链反应（RT-PCR）法检测β-catenin和细胞周期蛋白D1（Cyclin D1） mRNA表达,Western blot法检测Axin 1、β-catenin、磷酸化β-catenin（p-β-catenin）和Cyclin D1的表达.结果 不同浓度的XAV939对胃癌细胞株MKN-45细胞增殖的抑制率分别是22.5％、39.1％、52.5％、62.1％（P＜0.05）;不同浓度的XAV-939处理组的β-catenin、Cyclin D1 mRNA表达下降（分别为0.727±0.031、0.467±0.051、0.353±0.035、0.293±0.031,P ＜0.05;0.690 ±0.161、0.487±0.231、0.360±0.257、0.216±0.197,P ＜0.05）,β-catenin、Cyclin D1蛋白表达下降而Axin 1和p-β-catenin蛋白表达上升.结论 XAV-939可抑制以胃癌细胞株MKN-45的增殖,机制可能是通过抑制Wnt/β-catenin通路.

Objective To investigate the effects of XAV939 on proliferation and and Wnt/β-catenin signaling pathway of MKN-45 cells in vitro.Methods MKN-45 cells were cultured with different concentrations of XAV939.The inhibition of cell proliferation was detected by methyl thiazol tetrazolium （MTT）.The expression levels of β-catenin and cell cycle protein 1 （Cyclin D1） mRNA were detected by reverse transcription-polymerase chain reaction （RT-PCR）,and the protein expression of Axin 1,β-catenin,p-β-catenin and Cyclin D1 by Western blotting.Results Inhibitory rate of MKN-45 cells treated with XAV939 for 48 h was 22.5%,39.1%,52.5%,and 62.1% respectively （P 〈0.05）.Different concentrations of XAV939 could down-regulate β-catenin and Cyclin D1 mRNA level （0.727 ±0.031,0.467 ±0.051,0.353 ±0.035,0.293 ±0.031,P〈0.05; 0.690±0.161,0.487 ±0.231,0.360 ±0.257,0.2 1 6 ± 0.197,P 〈 0.05）,down-regulate β-catenin and Cyclin D1 protein levels,but significantly up-regulate Axin 1 and p-β-catenin expression levels.Conclusion XAV939 can inhibite proliferation of MKN-45 cells,which may be associated with down-regulation of Wnt/β-catenin signaling pathway.