摘要
目的：研究白藜芦醇苷（polydatin）在体外对乳腺癌细胞MCF-7增殖的作用,进一步探索其潜在的机制。方法：设定空白组及白藜芦醇苷（0.2,0.4,1.2,2.0,2.8 mg·L^-1）组,5-氟尿嘧啶组,采用噻唑蓝（MTT）比色法测定不同质量浓度白藜芦醇苷对MCF-7细胞的生长抑制率;应用细胞克隆集落形成实验验证白藜芦醇苷对MCF-7细胞增殖的抑制作用;通过实时荧光定量PCR（Real-time PCR）检测5-氟尿嘧啶（1.0μg·L^-1）和白藜芦醇苷（0.2,0.4,1.2 mg·L^-1）对DNA聚合酶δ催化亚基P125（P125）,P125编码基因（POLD1）,p53基因（p53）,细胞周期依赖性蛋白激酶抑制因子p21 mRNA表达水平的影响;应用蛋白质免疫印迹法（Western blot）检测白藜芦醇苷（0.2,0.4,1.2 mg·L^-1）对P125,p53,p21蛋白表达水平的影响。结果：与空白组比较,白藜芦醇苷（0.2,0.4,1.2,2.0,2.8 mg·L^-1）在体外对MCF-7细胞具有明显的抑制作用（P〈0.05,P〈0.01）,并且这种抑制作用呈浓度依赖性。与空白组比较,白藜芦醇苷（0.2,0.4,1.2 mg·L^-1）均能明显抑制细胞集落的形成（P〈0.05,P〈0.01）。与空白组比较,5-氟尿嘧啶组和白藜芦醇苷（0.4,1.2 mg·L^-1）能够显著抑制POLD1 mRNA表达水平,同时显著抑制P125 mRNA和蛋白表达水平,明显升高p53,p21 mRNA和蛋白表达水平（P〈0.05,P〈0.01）。与5-氟尿嘧啶组比较,白藜芦醇苷（1.2 mg·L^-1）对POLD1,P125,p53,p21 mRNA和蛋白表达水平影响均无显著差异。结论：白藜芦醇苷在体外能够显著抑制MCF-7细胞的增殖,其潜在机制可能与提高p53表达水平,抑制POLD1表达,促进p21表达有关。
Objective： To study the effect of polydatin on the proliferation of MCF-7 cells,in order to further explore its underlying mechanism.Method： MCF-7 cells were treated with polydatin at different doses（0,0.2,0.4,1.2,2.0,2.8 mg·L^-1）.The inhibitory rate of the cells was detected by methylthiazolydiphenyltetrazolium bromide（MTT） assay after treatment for 48 h.To further verify the inhibitory effect of polydatinon on the proliferation of MCF-7 cells,the cell colonies were detected by the colony formation assay after treatment with polydatin（0.2,0.4,1.2 mg·L^-1） for 10 days.After treatment with polydatin for 48 h,the effect of polydatin（0.2,0.4,1.2 mg·L^-1） on mRNA expressions of DNA polymerase δ catalytic subunit gene POLD1,P125,p53 and p21 were detected by Real-time PCR.Protein expressions of P125,p53 and p21 were detected by Western blot.Result： Polydatin at different doses（0.2,0.4,1.2,2.0,2.8 mg·L^-1） showed an inhibitory effect on the proliferation of MCF-7 cells in a concentration-dependent manner（P〈0.05,P〈0.01）.After administration for10 days,polydatin（0.2,0.4,1.2 mg·L^-1） significantly inhibited the formation of cell colonies,compared with control group（P〈0.05,P〈0.01）.After treatment for 48 h,mRNA expressions of POLD1 and P125 were significantly lower in polydatin（0.4,1.2 mg·L^-1） groups and 5-FU group than control group,while mRNA expressions of p53 and p21 were increased significantly（P〈0.05,P〈0.01）.After treatment for 48 h,protein expression of P125 was obviously lower in polydatin（0.4,1.2 mg·L^-1） groups and 5-FU group than control group,while protein expressions of p53 and p21 were increased significantly.The effect of polydatin on mRNA and protein expressions of PLOD1,P125,p53 and p21 in polydatin（1.2 mg·L^-1） group had no difference with 5-FU group.Conclusion： Polydatin shows a significantly inhibitory effect on the proliferation of MCF-7 cells.Its underlying mechanisms may be associated with the up-regulation of p53,the inhibit
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第4期143-148,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81360396)
广西壮族自治区卫生和计划生育委员会自筹经费科研课题项目(Z2015613)
广西医科大学青年科学基金项目(GXMUYSF201405)
