摘要
目的通过体内外实验,探讨穿心莲内酯(andrographolide,Andro)对三阴性乳腺癌细胞增殖、迁移、干细胞特性和上皮间质转化进程的影响及可能作用机制。方法利用磺酰罗丹明B(sulforhodamine B,SRB)比色法检测细胞活性;采用细胞划痕实验、流式技术和干细胞成球实验检测细胞迁移能力、干细胞增殖能力及CD44^(+)/CD24^(-/low)的比例;Western Blot实验检测NESTIN、NANOG、E-cadherin、Vimentin、N-cadherin、Fibronectin、p-YAP、YAP、ANKRD1蛋白的表达。采用小鼠移植瘤实验检测Andro对乳腺癌细胞体内增殖和小鼠致瘤性的作用。利用免疫组化实验检测各组肿瘤组织中YAP和ANKRD1的表达。结果体外实验结果表明:Andro可剂量依赖性降低MDA-MB-231和4T1细胞活性;与对照组比较,经Andro干预后的MDA-MB-231和4T1细胞的划痕愈合率降低,CD44^(+)/CD24^(-/low)比例下降,干细胞微球的体积减小和数量降低(P<0.05),NESTIN、NANOG、Vimentin、N-cadherin、Fibronectin、YAP和ANKRD1蛋白表达下降,而E-cadherin和p-YAP的表达上升。体内实验结果表明:与对照组相比,经Andro干预的小鼠肿瘤体积和肿瘤质量减小,肿瘤组织中YAP和ANKRD1的表达下降(P<0.01)。结论Andro可通过调控Hippo/YAP信号通路发挥抗三阴性乳腺癌的作用。
Objective To explore the effect of andrographolide on proliferation,migration,stem cell properties and epithelial-mesenchymal transition process of triple negative breast cancer cells and its mechanism through in vivo and in vitro experiments.Methods Sulforhodamine B method was used to detect the Cell viability.Migration,the proportions of CD44^(+)/CD24^(-/low) and the proliferation level of cancer stem cells were detected by wound healing assay,flow cytometry andmanmospheres assay,respectively.Western blot was used to detect the effects of andrographolide on the expression levels of YAP,ANKRD1,NESTIN,NANOG,E-cadherin,Vimentin,Ncadherin,Fibrone-Ctin YAP,ANKRD1,NESTIN,NANOG,E-cadherin,Vimentin,N-cadherin,Fibronectin and p-YAPin MDA-MB-231 and 4T1 cells.The effect of andrographolide on proliferation of breast cancer cells in vivo and the tumorigenicity of mice were examined through tumor transplantation experiment in mice.The expressions of YAP and ANKRD1 of tumor in mice was detected by immunohistochemistry.Results In vitro experiment results showed that cell viability of MDA-MB-231 and 4T1 cellstreated with andrographolide was decreased.in a dosedependent manner.Compared with the control group,the healing rate of cells,the proportions of CD44^(+)/CD24^(-/low) andthe volume and the number of stem cell microspheres in MDA-MB-231 and 4T1 cells were reduced(P<0.05).After andrographolide treatment,the expressions of YAP,ANKRD1,NESTIN,NANOG,Vimentin,N-cadherin and Fibronectin in MDA-MB-231 and 4T1 cells were down-regulated.Meanwhile,while the expression of E-cadherin andp-YAP were increased.In vivo experiment results showed that Compared with the control group,tumor volume and weight decreased and tumor inhibitory rate increased significantly after Andrographolide inter⁃vention(P<0.01).And the expression of YAP andANKRD1 of tumor in mice decreased.Conclusion Andrographolide may play a role against triple-negative breast cancer by regulating the Hippo/YAP pathway.
作者
黄翠霞
张雅倩
杨爱萍
刘芮含秋
路艳
HUANG Cuixia;ZHANG Yaqian;YANG Aiping;LIU Ruihanqiu;LU Yan(Research Center for Integrative Medicine,college of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)
出处
《实用医学杂志》
CAS
北大核心
2023年第16期2050-2056,共7页
The Journal of Practical Medicine
基金
广东省普通高校特色创新项目(编号:2019KTSCX028)。
关键词
三阴性乳腺癌
穿心莲内脂
YAP蛋白
上皮间质转化
triple negative breast cancer
andrographolide
yes-associatedprotein
epithelialmesenchymal transition
