槲皮黄酮调控Hedgehog通路抑制人卵巢癌细胞株A2780增殖、侵袭、迁移的体外实验  

Inhibitory Effect of Quercetin on Proliferation,Invasion and Migration of Human Ovarian Cancer Cell Line A2780 By Hedgehog Pathway in Vitro

作　　者：黄艳 徐锋[2] 梁博 刘思源 HUANG Yan;XU Feng;LIANG Bo;LIU Siyuan(Shenzhen University General Hospital,Shenzhen Guangdong China 518055;The First Hospital of Shijiazhuang,Shijiazhuang Hebei China 050011;Zhengzhou Railway Vocational and Technical College,Zhengzhou Henan China 451460)

机构地区：[1]深圳大学总医院,广东深圳518055 [2]石家庄市第一医院,河北石家庄050011 [3]郑州铁路职业技术学院,河南郑州451460

出　　处：《中医学报》2020年第9期1939-1945,共7页Acta Chinese Medicine

基　　金：国家科技部科技基础性工作专项基金项目(2017FY8361000)。

摘　　要：目的:通过体外实验研究分析槲皮黄酮通过调控Hedgehog通路对人卵巢癌细胞株A2780的增殖、侵袭和迁移的抑制作用。方法:将人卵巢癌细胞株A2780按照每孔1×10^5个接种到96孔板中,分别记作阴性对照组,阳性对照组,槲皮黄酮高剂量组、中剂量组、低剂量组,每组设置5个重复孔。阳性对照组培养基中加入阿霉素使其终浓度为5μmol·L^-1,槲皮黄酮高剂量组、中剂量组、低剂量组分别在培养基中加入槲皮黄酮使其终质量浓度为20、10、5μg·L^-1,培养48 h。通过四唑盐比色法分别在24 h和48 h检测A2780细胞增殖抑制率;通过Transwell小室法检验A2780细胞侵袭能力;通过细胞划痕实验检测A2780细胞迁移能力;实时荧光定量聚合酶链式反应检验Sonic Hedgehog(Shh)、Ptched1(Ptch1)、Smoothened(Smo)、神经胶质瘤相关癌基因同源蛋白1(Gli1)mRNA表达水平;通过蛋白免疫印迹检测Shh、Ptch1、Smo、Gli1蛋白表达水平。结果:阴性对照组A2780细胞在药物干预24 h和48 h细胞增殖抑制率比较,差异无统计学意义(P>0.05);其余4组48 h均高于24 h(P<0.05);两组间在24 h和48 h细胞增殖抑制率比较:槲皮黄酮高剂量组>阳性对照组/槲皮黄酮中剂量组>槲皮黄酮低剂量组>阴性对照组,阳性对照组和槲皮黄酮中剂量组比较,差异无统计学意义(P>0.05),其余各组间差异均有统计学意义(P<0.05)。药物干预48 h后A2780细胞侵袭活性和迁移能力比较:每两组比较,阴性对照组>槲皮黄酮低剂量组>阳性对照组/槲皮黄酮中剂量组>槲皮黄酮高剂量组,阳性对照组和槲皮黄酮中剂量组之间比较,差异无统计学意义(P>0.05),其余各组间比较,差异均有统计学意义(P<0.05)。药物干预48 h后Shh、Ptch1、Smo、Gli1 mRNA和蛋白表达水平检测比较:每两组间比较,阴性对照组>槲皮黄酮低剂量组>阳性对照组/槲皮黄酮中剂量组>槲皮黄酮高剂量组,阳性对照组和槲皮黄酮中剂量组之间�Objective:To analyze the inhibitory effect of quercetin on proliferation,invasion and migration of human ovarian cancer cell line A2780 by regulating Hedgehog pathway though in vitro experiments.Method:Human ovarian cancer cell line A2780 was inoculated into 96-well plate according to 1×10^5 wells,which were divided into negative control group,positive control group,high dose group,medium dose group and low dose group of quercitrin,and each group was provided with 5 repeated wells.In the positive control group,adriamycin was added to the medium to make the final concentration reach 5μmol·L^-1,and the high-dose,medium-dose and low-dose groups were added with quercetin to make the final concentration reach 20μg·L^-1,10μg·L^-1 and 5μg·L^-1,respectively.The proliferation inhibition rate of A2780 cells was detected by MTT assay at 24 h and 48 h,respectively;the invasion ability of A2780 cells was tested by Transwell chamber method;the migration ability of A2780 cells was detected by cell scratch test;the mRNA expression levels of Sonic Hedgehog(Shh),Ptched1(Ptch1),Smoothened(Smo)and glioma-associated oncogene homologous protein 1(Gli1)were detected by real-time fluorescence quantitative polymerase chain reaction(RTqPCR);The expression levels of Shh,Ptch1,Smo and Gli1 were detected by western blot(WB).Result:There was no significant difference in the inhibition rate of A2780 cells in negative control group between 24h and 48h after drug intervention(P>0.05).In the other four groups,48 h was higher than 24 h(P<0.05).Comparing the inhibition rate of cell proliferation between the two groups at 24 h and 48 h,the results showed that:high-dose quercetin group>positive control group/middle-dose quercetin group>low-dose quercetin group>negative control group.There was no significant difference between the positive control group and middle-dose quercetin group(P>0.05),but there were significant differences among other groups(P<0.05).Comparison of invasion activity and migration ability of A2780 cells after 48 h of dru

关 键 词：槲皮黄酮 卵巢癌 细胞增殖 细胞侵袭 细胞迁移 HEDGEHOG通路 

分 类 号：R285.5[医药卫生—中药学]