槲皮素通过靶向GAS5/Notch1信号通路促进乳腺癌细胞凋亡的研究  

Quercetin promotes apoptosis of breast cancer cells by targeting GAS5/Notch1 signaling pathway

作　　者：蒋多晨 金乐 陈洪晓 张蕾[2] 陈昭琳 居靖[1] JIANG Duo-chen;JIN Le;CHEN Hong-xiao;ZHANG Lei;CHEN Zhao-lin;JU Jing(Anqing Hospital Affiliated to Anhui Medical University,Anqing,Anhui 246003,China;Dept of Pharmacy,the First Affiliated Hospital of USTC,Division of Life Sciences and Medicine,Anhui Provincial Hospital,Hefei 230001,China)

机构地区：[1]安徽医科大学附属安庆医院,安徽安庆246003 [2]中国科学技术大学附属第一医院(安徽省立医院)药剂科,安徽合肥230001

出　　处：《中国药理学通报》2021年第5期637-644,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81602344);中国博士后科学基金面上资助项目(No 2019M662207);安徽省重点研究与开发计划项目(No 201904a07020092)。

摘　　要：目的探讨槲皮素对乳腺癌细胞促凋亡作用的可能机制。方法用槲皮素处理乳腺癌MCF-7细胞后,采用MTT法检测细胞活力;平板克隆实验检测细胞增殖克隆能力;Annexin V-FITC/PI双染法检测细胞凋亡;同时,分别采用槲皮素(40、80、160μmol·L^(-1))、GAS5小干扰RNA(siGAS5)、槲皮素(80μmol·L^(-1))协同siGAS5处理细胞后,qRT-PCR法和Western blot法检测GAS5、Notch1、Jagged1、Hes1、Bcl-2、Bax、Caspase-3的表达。结果槲皮素(40、80、160μmol·L^(-1))处理MCF-7细胞后,发现40μmol·L^(-1)槲皮素可明显抑制增殖,而80、160μmol·L^(-1)槲皮素不仅明显抑制增殖且诱导凋亡;槲皮素(40、80、160μmol·L^(-1))可上调GAS5、Bax和Caspase-3的表达,下调Notch1、Jagged1、Hes1和Bcl-2的表达;细胞转染siGAS5后,与sncRNA组相比,GAS5、Bax和Caspase-3表达下调,Notch1、Jagged1、Hes1和Bcl-2表达上调;当siGAS5与槲皮素(80μmol·L^(-1))共处理细胞后,与sncRNA加槲皮素对照组相比,GAS5、Bax和Caspase-3表达下调,Notch1、Jagged1、Hes1和Bcl-2上调。结论槲皮素通过靶向GAS5/Notch1信号通路促进乳腺癌细胞凋亡。Aim To investigate the possible mechanism of quercetin in promoting apoptosis of breast cancer cells.Methods Breast cancer cells MCF-7 were treated with quercetin.Cell viability was detected by MTT assay,cell proliferation and cloning ability were measured by plate colony formation assay,and cell apoptosis was examined by Annexin V-FITC/PI double staining assay.Meanwhile,MCF-7 cells were treated with quercetin(40,80,160μmol·L^(-1)),small interfering RNA of GAS5(siGAS5)and quercetin(80μmol·L^(-1))in combination with siGAS5,respectively.The expressions of GAS5,Notch1,Jagged1,Hes1,Bcl-2,Bax and Caspase3 in MCF-7 cells were analysed by qRT-PCR and Western blot.Results After treatment of MCF-7 cells with quercetin,it was found that quercetin at concentration of 40μmol·L^(-1) significantly inhibited the proliferation of MCF-7 cells,and quercetin at concentration of 80μmol·L^(-1) or 160μmol·L^(-1) not only inhibited proliferation but also induced apoptosis.Quercetin(40,80,160μmol·L^(-1))could up-regulate the expressions of GAS5,Bax,Caspase3,and down-regulate the expressions of Notch1,Jagged1,Hes1,Bcl-2 in MCF-7 cells.In comparison with sncRNA group,the expressions of GAS5,Bax and Caspase3 were down-regulated and the expressions of Notch1,Jagged1,Hes1 and Bcl-2 were up-regulated after transfection with siGAS5.And when MCF-7 cells transfected with siGAS5 and then cotreated with 80μmol·L^(-1) quercetin,we found that compared to quercetin combined with sncRNA,the expressions of Notch1,Jagged1,Hes1 and Bcl-2 were up-regulated,while GAS5,Bax and Caspase3 were down-regulated.Conclusion Quercetin promotes apoptosis of breast cancer cells by targeting GAS5/Notch1 signaling pathway.

关 键 词：槲皮素 乳腺癌 长链非编码RNA GAS5 Notch1信号通路 细胞凋亡 靶向 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R342.2[医药卫生—基础医学]