槲皮素对人黑素瘤A375细胞增生和凋亡的影响  

Effects of quercetin on the proliferation and apoptosis of A375 human melanoma cells

作　　者：卫艳萍[1] 史四季[1] 张如幸 刘秀琴[1] 赵林栋[1] 

机构地区：[1]焦作市人民医院皮肤性病科,焦作454002

出　　处：《实用皮肤病学杂志》2016年第3期171-174,共4页Journal of Practical Dermatology

摘　　要：目的探讨槲皮素对体外培养人黑素瘤A375细胞增生和凋亡的影响。方法 CCK8法检测0、20、40、80、160μmol/L浓度槲皮素作用于A375细胞24、48和72 h后细胞增生水平。用0、20、40、80、160μmol/L浓度槲皮素作用24 h后,以Annexin-V/PI法观察A375细胞凋亡,分别测定Caspase 3、Caspase 8和Caspase 9的活性,并以蛋白印迹法(Western blot)检测p53,Bax及Bcl-2的基因蛋白水平。结果与空白对照组比较,20、40、80、160μmol/L槲皮素作用24、48和72 h后均对A375细胞的增生有抑制作用,且有时间、浓度依赖性。0、20、40、80、160μmol/L槲皮素在作用24 h时,A375细胞早期凋亡率由(3.02±0.49)%分别上升至(11.20±1.68)%、(12.44±1.68)%、(24.55±0.58)%和(47.68±0.79)%,各组与对照组相比差异均有统计学意义(P<0.05或<0.01),80、160μmol/L作用组的Caspase 3,Caspase8,Caspase 9活性升高,且与对照组比较,差异均有统计学意义(P<0.05或<0.01)。p53及Bax的蛋白水平随药物浓度的增加而升高,Bcl-2的蛋白水平随药物浓度增加而降低。结论槲皮素对黑素瘤A375细胞具有抑制增生和诱导凋亡的作用,其作用机制可能与调节p53及Bcl-2/Bax基因有关。Objective To study the effect of quercetin on the proliferation and apoptosis of A375 human melanoma cells.Methods Cell counting kit-8（CCK8） was used to evaluate the proliferative activity of A375 cells which treated with quercetin of 0,20,40,80,160μmol/L for 24 h,48 h and 72 h respectively.Subsequently,A375 cells that treated with the same method underwent double staining with annexin V and propidium iodide for the detection of cell apoptosis,and fluorometric assay for the estimation of Caspase 3,Caspase 8 and Caspase 9 activity,and Western blot for the determination of the levels of p53,Bax and Bcl-2 proteins.Results Within the investigated concentration and time ranges,quercetin significantly inhibited the proliferative activity of A375 cells in a concentrationdependent and time-dependent manner.Compared with blank groups,a significant increase was observed in the early apoptosis rate in A375 cells treated with quercetin of 0,20,40,80,160 μmol/L for 24 h（11.20±1.68）%,（12.44±1.68）%,（24.55±0.58）% and（47.68±0.79）% vs（3.02±0.49）%,（P〈0.05 or 0.01）,as well as in the activity of Caspase 3,8 and 9 in A375 cells treated with quercetin of 80,160μmol/L for 24 h（P〈0.05 or 0.01）.After 24 h treatment,the protein levels of p53 and Bax were elevated,but Bcl-2 was decreased in A375 cells with the increase of quercetin concentration.Conclusion Quercetin can inhibit the proliferation and induce the apoptosis of A375 cells,and the apoptosis-inducing effect may occur through modulating the express of p53 and Bcl-2/Bax genes.

关 键 词：黑素瘤 槲皮素 凋亡 增生 

分 类 号：R739.5[医药卫生—肿瘤]