摘要
目的探讨白花蛇舌草总黄酮(FOD)对MDA-MB-231乳腺癌细胞增殖、凋亡及干性的影响。方法培养MDA-MB-231细胞系,分别用0、100、200、400μg/mL FOD作用MDA-MB-231乳腺癌细胞不同时间(24、48、72 h),CCK8(Cell Counting Kit-8)法检测其对细胞增殖的影响;平板克隆试验检测各组MDA-MB-231细胞增殖能力变化;Annexin V-PE/7-AAD法检测各组MDA-MB-231细胞凋亡比例,并用Western blotting法检测Bcl2、Bax及凋亡指标cleaved-caspae3等凋亡通路蛋白的表达,流式细胞检测CD44+/CD24-乳腺癌细胞干性,并用Western blotting法检测干性肿瘤指标Nanog、Sox2、Oct4的表达量。结果CCK8实验显示,400μg/mL FOD作用72 h(FOD组)后细胞增殖较阴性对照组(DMSO组)受到明显抑制(P<0.05);平板克隆试验显示,药物干预组细胞增殖下降(P<0.05);经400μg/mL FOD处理的MDA-MB-231干性标志物Nanog、Oct4、Sox2表达更低(P<0.05)。流式细胞仪检测FOD组细胞凋亡比例较阴性对照组明显升高(P<0.05),抗凋亡蛋白Bcl-2表达降低,而促凋亡蛋白Bax及凋亡指标cleaved-caspae3的蛋白表达升高,FOD处理后乳腺癌细胞增殖、凋亡及干性改变,通路蛋白Akt及GSK3β磷酸化受到抑制,β-catenin表达降低。结论FOD可明显抑制MDA-MB-231乳腺癌细胞增殖及干性,促进细胞凋亡。
Objective To investigate the effect of total flavone of oldenlandia diffusa(FOD)on the stemness,proliferation and apoptosis of breast cancer(BC)stem cells sorted from MDA-MB-231.Methods Human BC cell lines MDA-MB-231 was cultured in vitro;MDA-MB-231 was stimulated by different concentrations(0μg/mL,100μg/mL,200μg/mL and 400μg/mL)of FOD for different time(24 h,48 h and 72 h).CCK8 and plate cell cloning assay were used to detect the effect of FOD on MDA-MB-231 proliferation;CD44+/CD24-MDA-MB-231 cell line were tested by flow cytometry and stem cell markers such as Nanog,Oct4 and Sox2 were tested by Western blotting;Annexin V-PE/7-AAD was used to detect the effect of FOD on MDA-MB-231 apoptosis and Bcl2,cleaved-caspase3 and Bax were tested by Western blotting.Results Cell proliferation of MDA-MB-231 was significantly inhibited by FOD,with the significant suppression at concentrations of 400μg/mL for 72 h compared with negative control group(P<0.05).The apoptosis rate was significantly upregulated than the negative control group(P<0.05).The protein expression of Bcl2 decreased while Bax and cleaved-caspae3 increased,and stemness markers such as Nanog,Sox2 and Oct4 decreased in FOD-treated cells.Moverover,Akt-GSK3β-β-catenin axis was inhibited in FOD-treated cells.Conclusion FOD could significantly inhibit the stemness and proliferation and promote the apoptosis of MDA-MB-231.
作者
姚博文
李亚昭
李静宇
李超逸
鲁叶
马婕群
张彦兵
YAO Bowen;LI Yazhao;LI Jingyu;LI Chaoyi;LU Ye;MA Jiequn;ZHANG Yanbing(Department of Hepatobiliary Surgery,The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061;Center for Tranlational Medicine,The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061;Zonglian College,Xi’an Jiaotong University,Xi’an 710061;Department of Oncology,Shaanxi Provincial Cancer Hospital,Xi’an 710061,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2023年第6期880-885,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
陕西省自然科学基础研究计划(No.2023-JC-YB-680)
西安交通大学第一附属医院科研发展基金国科金青年储备项目(No.2021QN-04)
陕西省重点研发计划基金资助项目(No.2021SF-220)。
关键词
乳腺癌
干细胞
白花蛇舌草总黄酮
增殖
凋亡
breast cancer
stem cell
total flavone of oldenlandia diffusa(FOD)
proliferation
apoptosis
作者简介
通信作者:张彦兵,副主任医师。E-mail:zhang_yb007@163.com。
