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目的研究Quercetin对结肠癌LOVO细胞株TGF-β1/smad3/c-myc信号通路的影响,并探讨其抑制LOVO细胞增殖的可能机制。方法采用5、10、20、40、80、160μmol/L的Quercetin处理结肠癌LOVO细胞48 h,以未经Quercetin处理的LOVO细胞作为对照组,采用MTT法检测Quercetin对细胞增殖活力的影响。以5 ng/ml的TGF-β1刺激1周的LOVO细胞为细胞模型,设对照组(C组)、TGF-β1组(T组)、TGF-β1+Quercetin组(TQ组)、Quercetin组(Q组)。C组为正常的LOVO细胞;T组为用5 ng/ml的TGF-β1刺激1周的LOVO细胞;TQ组为以5 ng/ml的TGF-β1刺激1周后,再经20μmol/L Quercetin处理72 h的LOVO细胞;Q组为经20μmol/L Quercetin处理72 h的LOVO细胞。分别采用平板克隆试验、免疫组化SP法、RT-PCR法、Western blot法分析Quercetin和TGF-β1对LOVO细胞克隆形成能力、smad3及c-myc表达的影响。结果与对照组相比,不同浓度Quercetin组可明显抑制LOVO细胞的增殖,且呈剂量依赖性,IC50值约为40μmol/L。TGF-β1能明显促进LOVO细胞的增殖及smad3、c-myc的表达(P<0.05);Quercetin能明显抑制LOVO细胞的增殖及smad3、c-myc的表达(P<0.05);与Q组相比,TQ组细胞增殖能力及c-myc的表达明显降低(P<0.05)。结论 Quercetin通过抑制TGF-β1/smad3信号通路下调靶基因c-myc的表达,并具有抑制LOVO细胞增殖的作用。
Objective To investigate the effect of Quercetin on TGF-β1 / smad3 / c-myc signaling pathway in LOVO cell lines and to explore its possible mechanism of inhibiting the proliferation of LOVO cells. Methods LOVO cells were treated with Quercetin at 5, 10, 20, 40, 160 and 160μmol / L for 48 h. Quercetin-treated LOVO cells were used as control group. The effect of Quercetin on cell proliferation was measured by MTT assay. The cells in the control group (group C), TGF-β1 group (group T), TGF-β1 + Quercetin group (group TQ) and Quercetin group Q group). Group C was normal LOVO cells; Group T was LOVO cells stimulated with 5 ng / ml TGF-β1 for 1 week; Group TQ was stimulated with 5 ng / ml TGF-β1 for 1 week and then treated with 20 μmol / L Quercetin LOVO cells were treated for 72 h and Q cells were LOVO cells treated with 20 μmol / L Quercetin for 72 h. The effects of Quercetin and TGF-β1 on the clonality, smad3 and c-myc expression in LOVO cells were analyzed by plate clone assay, immunohistochemical SP method, RT-PCR and Western blot respectively. Results Compared with the control group, different concentrations of Quercetin group could significantly inhibit the proliferation of LOVO cells in a dose-dependent manner with IC50 of about 40μmol / L. TGF-β1 could obviously promote the proliferation of LOVO cells and the expression of smad3 and c-myc (P <0.05). Quercetin could significantly inhibit the proliferation and the expression of smad3 and c-myc in LOVO cells (P <0.05) The cell proliferation and c-myc expression in TQ group were significantly lower than those in TQ group (P <0.05). Conclusion Quercetin can down-regulate the expression of c-myc target gene by inhibiting TGF-β1 / smad3 signaling pathway and inhibit the proliferation of LOVO cells.