槲皮素对宫颈癌细胞C33A增殖和凋亡的影响

目的 观察槲皮素对人宫颈癌细胞C33A增殖和凋亡的影响,初步探讨其相关作用机制.方法 用不同浓度槲皮素(空白对照、20、40、80 μmol/L),顺铂(空白对照、5、10、15、20 μg/mL)以及两者联合(槲皮素40 μmol/L+顺铂10 μg/mL)分别作用于C33A细胞24 h和48 h后,噻唑蓝(MTT法)检测细胞活力的变化;流式细胞术检测槲皮素对C33A细胞周期的影响;Western blot检测槲皮素对C33A细胞Cyclin D1、Caspase-3和Caspase-9蛋白表达的影响.结果 MTT结果显示,经过槲皮素处理24 h后,各组细胞活力分别为86.92 ±3.953)%(20 μmol/L组)、(66.54 ±3.932)%(40 μmol/L组)和(52.21 ±2.970)%(80 μmol/L组),均低于空白对照组的(98.35 ±1.230)%;经过槲皮素处理48 h后,各组细胞活力分别为(65.19 ±7.071)%(20 μmol/L组)、(47.04 ±8.881)%(40 μmol/L组)和(29.71 ±6.505)%(80 μmol/L组),均低于空白对照组的(96.97 ±1.788)%.;此外,槲皮素40 μmol/L+顺铂10 μg/mL联合作用于C33A细胞24h后,细胞活力下降为(31.12 ±2.835)%;48 h后,细胞活力下降为(17.86 ± 3.182)%,同空白对照组相比均出现了明显的下降,(31.12 ±2.835)%;48 h后,细胞活力下降为(17.86 ±3.182)%(P<0.05).细胞周期检测结果显示,槲皮素可增加C33A细胞G0/G1期数量,减少S期数量.Western blot 结果显示,槲皮素可下调C33A细胞Cyclin D1蛋白的表达,还可上调Caspase-3和Caspase-9蛋白的表达.结论 槲皮素通过下调Cyclin D1蛋白的表达可以抑制C33A细胞的活力和增殖,槲皮素通过上调Caspase-3和Caspase-9蛋白的表达,可以诱导C33A细胞的凋亡.

Effect of quercetin on proliferation and apoptosis of cervical cancer cell C33A

Objective To investigate the effect of quercetin on proliferation and apoptosis of human cervical cancer cell line C 33A and its mechanisms.Methods C33A was treated with quercetin alone(blank control,20,40,80 μmol/L),combined with cisplatin(querce-tin 40 μmol/L+cisplatin 10 μg/mL)or cisplatin alone(blank control,5,10,15,20 μg/mL)at different time points(24 h and 48 h).MTT assay was used to detect cell viability;flow cytometry was employed to detect cell cycle in quercetin treated C 33A cells;Western blotting was applied to detect the expression of Cyclin D 1,Caspase-3 and Caspase-9 in quercetin treated C33A cells.Results By treatment with querce-tin for 24 h,cell vitality was(86.92 ±3.953)%(20 μmol/L),(66.54 ±3.932)%(40 μmol/L)and(52.21 ±2.970)%(80 μmol/L);by treatment with quercetin for 48h,the cell vitality was(65.19 ±7.071)%(20 μmol/L),(47.04 ±8.881)%(40 μmol/L)and(29.71 ± 6.505)%(80 μmol/L),compared with(96.97 ±1.788)%in blank control group(P<0.05);moreover,after treatment with quercetin(40 μmol/L)+cisplatin(10 μg/mL),cell vitality decreased to(31.12 ±2.835)%(24 h)and(17.86 ±3.182)%(48 h)(P<0.05). Quercetin could increase the number of G 0/G1 cells and reduce the number of S cells in C 33A cells.Quercetin could down -regulate the expression of Cyclin D1 and up-regulate the expression of Caspase -3 and Caspase-9 in C33A cells.Conclusion Quercetin inhibits cell vitality and cell proliferation by down -regulating the expression of Cyclin D1 in C33A cells, and it may also induce cell apoptosis through up-regulating the expression of Caspase -3 and Caspase-9.