大黄素甲醚-8-O-β-D-葡萄糖苷对卵巢癌细胞周期影响机制探讨  

Effect of physcion-8-O-β-D-glucoside on ovarian cancer cell cycle and the underlying mechanism

作　　者：贺媛琪 李晓晓 张世红[1] 张建忠[1] 崔唯唯 陶晶岩 HE Yuan-qi;LI Xiao-xiao;ZHANG Shi-hong;ZHANG Jian-zhong;CUI Wei-wei;TAO Jing-yan(Department of Gynaecology and Obstetrics,Weihai Municipal Hospital,Weihai 264200,P.R.China)

机构地区：[1]威海市立医院妇产科,山东威海264200

出　　处：《中华肿瘤防治杂志》2020年第21期1703-1709,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省医药卫生科技发展计划(2017WSA10016)。

摘　　要：目的大黄素甲醚-8-O-β-D-葡萄糖苷(physcion 8-O-beta-D-monoglucoside,PG)作为天然产物已被证实具有抗肿瘤作用。本研究探讨PG对卵巢癌SK-OV-3细胞周期的影响并其可能的机制。方法卵巢癌SK-OV-3细胞作为研究对象,5、10、50、100和200μg/mL PG处理SK-OV-3细胞24h,结晶紫染色法观察细胞增殖能力,MTT法检测细胞活力并计算IC50。进一步分组为空白对照组、DMSO组、PG组、KU60019组和PG+KU60019组,PG(53.5μg/mL)处理细胞24h,KU60019(3μmol/L)预处理细胞4h,流式细胞术检测细胞周期分布,蛋白质印迹法检测p53、Cyclin E和CDK2蛋白表达变化。SPSS 17.0对数据进行统计学分析。结果与溶剂对照相比,10、50、100和200μg/mL PG处理后细胞相对增殖率明显降低,且呈剂量依赖性,F=140.30,P<0.001;与溶剂对照相比,10、50、100和200μg/mL PG处理后细胞细胞活力明显降低,且呈剂量依赖性,F=125.00,P<0.001。PG对SK-OV-3细胞的IC50为(53.5±4.2)μg/mL。KU60019可逆转PG(53.5μg/mL)诱导的细胞相对增殖率降低(t=14.02,P<0.001)。PG可诱导G1期细胞比例升高(F=5.84,P=0.008);而KU60019能够减弱PG对细胞周期的影响(F=18.90,P=0.025)。PG能诱导p53蛋白表达上调(F=173.50,P<0.001),同时诱导Cyclin E表达(F=42.65,P<0.001)和CDK2蛋白表达下调(F=43.72,P<0.001);而KU60019能够抑制PG诱导的p53蛋白表达上调(F=173.50,P<0.001),Cyclin E蛋白表达(F=42.65,P<0.001)和CDK2蛋白表达下调(F=43.72,P<0.001)。结论PG能抑制卵巢癌SK-OV-3细胞增殖并诱导G1期阻滞,该作用机制可能与增强ATM-P53信号通路转导进而抑制Cyclin E和CDK2蛋白表达有关。OBJECTIVE The anti-tumor effect of Physcion 8-O-beta-D-monoglucoside(PG),a natural product,has been proved.This study aimed to investigate influences of PG on SK-OV-3 cell cycle of ovarian cancer and possible influencing mechanism.METHODS Using ovarian cancer SK-OV-3 cells as research objects.SK-OV-3 cells were treated by different concentrations of PG(5,10,50,100 and 200μg/ml)for 24 h.Cell proliferative capacity was observed by crystal violet staining.Cell viability was detected by MTT method and IC50 was calculated.It was further divided into blank control group,DMSO group,PG group,KU60019 group and PG+KU60019 group.PG(53.5μg/ml)treated cells for 24 h,KU60019(3μmol/L)pretreated cells for 4 h.Cell cycle distributions were detected by flow cytometry.Changes of expressions of p53,Cyclin E and CDK2 protein were tested by protein western blots.RESULTS The relative cell proliferation rate of different experimental groups are significantly lower than that of the solvent control group.This reflects dose dependence of relative cell proliferation rate(F=140.30,P<0.001).The cell viability of different experimental groups are significantly lower than that of the solvent control group.This reflects dose dependence of cell viability(F=125.00,P<0.001).IC50 value of PG to SK-OV-3 cell is(53.5±4.2)μg/ml.KU60019 can reverse reduction of relative proliferation rate induced by PG(53.5μg/ml;t=14.02,P<0.001).PG can bring a increase of proportion of G1-phase cells(P=0.008),while KU60019 can weaken influences of PG on cell cycle(F=5.84,P=0.025).PG could induce overexpression of p53 protein(F=173.50,P<0.001),but downregulate expressions of Cyclin E(F=42.65,P<0.001)and CDK2(F=43.72,P<0.001).Moreover,KU60019 could inhibit influences of PG on expressions of p53(F=173.50,P<0.001),Cyclin E(F=42.65,P<0.001)and CDK2 protein(F=43.72,P<0.001).CONCLUSIONS PG can strengthen transduction of ATM-P53 signal pathway and thereby inhibit expressions of Cyclin E and CDK2.Therefore,PG can inhibit proliferation and induce blocking of G1-phase SK-OV-

关 键 词：大黄素甲醚-8-O-β-葡萄糖苷 卵巢癌 细胞周期 细胞周期关卡激酶ATM G1期 

分 类 号：R737.31[医药卫生—肿瘤]