摘要
目的:探讨槲皮素对人结肠癌细胞SW480增殖的影响及对细胞Wnt/β-catenin信号转导的调控作用。方法:以不同浓度的槲皮素处理SW480细胞,采用MTT比色法分析槲皮素对细胞增殖的抑制作用,利用报告基因方法研究槲皮素对Wnt/β-catenin信号转导的影响,用RT-PCR及蛋白质印迹法观察槲皮素对Wnt/β-catenin信号转导通路的下游关键因子Cyclin D1和Survivin转录水平和蛋白表达水平的影响。结果:(40～160)×10-6mol/L的槲皮素可明显抑制SW480细胞的增殖,P<0.01,抑制作用呈剂量依赖性和时间依赖性;(10～160)×10-6mol/L的槲皮素可明显抑制SW480细胞TCF-4报告基因TOPflash活性,P<0.05,抑制作用呈剂量依赖性。槲皮素可显著下调β-catenin/TCF下游靶基因CyclinD1和Survivin的mRNA表达和蛋白表达水平,呈剂量依赖性。结论:槲皮素可抑制人结肠癌细胞SW480的增殖,其抗肿瘤机制可能与其抑制Wnt/β-catenin信号转导通路有关。
OBJECTIVE： To investigate the relationship between the anti tumor activity of quercetin and the inhibition of Wnt/β catenin signaling pathway. METHODS： SW480 cells were incubated with quercetin at several concentrations and for different time. The MTT colorimetric assay was used to examine the effect of quercetin on the proliferation of SW480 cells. Reporter gene plasmid TOPflash or FOPflash was cotransfected into SW480 cells together with pRL-null plasmid, and the reporter gene assay was performed to detect the changes of the transcriptional activity of β-catenin / TCF in SW480 cells after quercetin was added. RT-PCR and Western blot analysis were performed to examine the transcriptional level and protein expression level of β-catenin target genes, Cyclin D1 and Survivin. RESULTS： The proliferation of SW480 cells was inhibited significantly by quercetin within the concentrations from 40 to 160 μmol/L, P〈0.01, showing a dose and time-dependent manner. The TCF-4 reporter gene activity in SW480 cells was inhibited significantly by quercetin within the concentrations from 10 to 160 μmol/L, P〈0.05, and mRNA and the protein expression of β-catenin/TCF target genes, Cyclin D1 and Survivin in SW480 cells were also down regulated markedly by quercetin, in a dose-ependent manner. CONCLUSION： Quercetin can inhibit the proliferation of SW480 cells in vitro and the anti-tumor mechanism may be related to the inhibition of Wnt/β-catenin signaling path＇way.
出处
《中华肿瘤防治杂志》
CAS
2008年第21期1632-1635,共4页
Chinese Journal of Cancer Prevention and Treatment
