鸡血藤总黄酮对人乳腺癌细胞株MCF-7增殖凋亡的影响及对Wnt/β-catenin通路的调控作用  

Effect of Total Flavonoids from Caulis Spatholepis on Human Breast Cancer Cell Line MCF-7 and Its Regulation of Wnt/β-catenin Pathway

作　　者：王兰英[1] 孙吉利 苏家茹[1] WANG Lanying;SUN Jili;SU Jiaru(Qingdao Municipal Hospital,Qingdao Shandong China 266011)

机构地区：[1]青岛市市立医院,山东青岛266011

出　　处：《中医学报》2021年第7期1512-1518,共7页Acta Chinese Medicine

基　　金：青岛市医药科研指导计划项目(2012-WSZD049)。

摘　　要：目的:研究鸡血藤总黄酮通过调控Wnt/β-catenin通路对人乳腺癌细胞株MCF-7增殖、迁移、凋亡等的影响。方法:将人乳腺癌细胞株MCF-7按每孔1×10~5个细胞接种到96孔板中,并分为阴性对照组、阳性对照组、鸡血藤总黄酮高、中、低剂量组共5组,培养48 h。MTT法检测MCF-7细胞增殖抑制率;TUNEL法检测MCF-7细胞凋亡指数;Transwell实验观察MCF-7细胞侵袭能力;细胞划痕实验观察MCF-7细胞迁移能力;RT-qPCR检测MCF-7细胞中Wnt1、β-catenin、CyclinD1、c-Myc及MMP-7 mRNA表达水平;Western Blot检测MCF-7细胞中Wnt1、β-catenin、CyclinD1、c-Myc和MMP-7蛋白表达水平及β-catenin磷酸化水平。结果:鸡血藤总黄酮显著抑制MCF-7细胞的增殖、侵袭及迁移(P<0.05),显著促进MCF-7细胞的凋亡(P<0.05);鸡血藤总黄酮显著下调Wnt1、β-catenin、cyclinD1、c-Myc和MMP-7的mRNA表达水平(P<0.05);鸡血藤总黄酮显著上调β-catenin磷酸化水平(P<0.05),显著下调Wnt1、β-catenin、cyclinD1、c-Myc和MMP-7蛋白的表达水平(P<0.05),且具有明显的剂量依赖性。结论:鸡血藤总黄酮可抑制人乳腺癌细胞MCF-7增殖、侵袭和转移能力,促进MCF-7细胞凋亡,机制可能与其调控Wnt/β-catenin信号通路有关。Objective:To study the effect of total flavonoids from caulis spatholepis on Wnt/β-catenin pathway on proliferation,migration and apoptosis of human breast cancer cell line MCF-7.Methods:The human breast cancer cell line MCF-7 was inoculated into a 96-well plate at 1×10~5 cells per well and divided into 5 groups:negative control group,positive control group,high,medium and low dose groups of total flavonoids from caulis spatholepis.The cells were cultured for 48 h.MTT method was used to detect the proliferation inhibition rate of MCF-7 cells;TUNEL method was used to detect the apoptosis index of MCF-7 cells;Transwell experiment was used to observe the invasion ability of MCF-7 cells;the migration ability of MCF-7 cells was observed by scratch test;RT-qPCR was used to detect the mRNA expression levels of Wnt1,β-catenin,CyclinD1,c-Myc and MMP-7 in MCF-7 cells;Western Blot was used to detect Wnt1,β-catenin,CyclinD1,c-Myc and MMP-7 protein expression levels andβ-catenin phosphorylation levels in MCF-7 cells.Results:The total flavonoids from caulis spatholepis significantly inhibited the proliferation,invasion and migration of MCF-7 cells(P<0.05),and significantly promoted the apoptosis of MCF-7 cells(P<0.05);the total flavonoids from caulis spatholepis significantly down-regulated Wnt1,β-catenin,cyclinD1,c-Myc and MMP-7 mRNA expression levels(P<0.05);the total flavonoids from caulis spatholepis significantly up-regulate the phosphorylation level ofβ-catenin(P<0.05),and significantly down-regulated the expression levels of Wnt1,β-catenin,cyclinD1,c-Myc and MMP-7 protein(P<0.05),and had a significant dose-dependent.Conclusion:The total flavonoids from caulis spatholepis can inhibit the proliferation,invasion and metastasis of human breast cancer cells MCF-7,and promote MCF-7 cell apoptosis.The mechanism may be related to the regulation of Wnt/β-catenin signaling pathway.

关 键 词：鸡血藤总黄酮 乳腺癌 增殖 凋亡 侵袭 迁移 WNT/Β-CATENIN通路 

分 类 号：R285.5[医药卫生—中药学]