基于Hedgehog通路探讨理肺消积丸对NSCLC A549细胞周期、增殖和迁移的影响
Exploring the Effects of Lifei-Xiaoji Pills on Cycle,Proliferation and Migration of NSCLC A549 Cell Based on Hedgehog Pathway
刘素晓 1张丛丛 2孔德琪 2黄慧敏 2毛静 3王元元 4刘学芳 3冯素香 3李亚1
作者信息
- 1. 河南中医药大学第一附属医院 郑州 450000;河南中医药大学呼吸疾病中医药防治省部共建协同创新中心 河南省中医药防治呼吸病重点实验室 郑州 450046
- 2. 河南中医药大学第一临床医学院 郑州 450046
- 3. 河南中医药大学呼吸疾病中医药防治省部共建协同创新中心 河南省中医药防治呼吸病重点实验室 郑州 450046
- 4. 河南中医药大学第一附属医院 郑州 450000
- 折叠
摘要
目的 探讨理肺消积丸对A549细胞的影响作用及对Hedgehog通路的调控作用.方法 采用MTT实验筛选理肺消积丸含药血清对A549细胞最佳干预浓度后,将细胞分为6组:对照组、理肺消积含药血清组、Hedgehog通路抑制剂组(Vismodegib组)、顺铂组、理肺消积含药血清+Vismodegib组、理肺消积含药血清+顺铂组.不同干预后,分别采用CCK-8实验检测细胞增殖能力,采用克隆形成实验观察细胞克隆形成能力,采用Transwell实验观察A549细胞迁移能力,采用流式细胞术观察细胞周期比例的变化,采用qRT-PCR和Western blot技术检测Hedgehog通路中Gli1、Ptch1、Smo和CyclinD1 mRNA和蛋白表达的变化.结果 浓度为10%的理肺消积丸含药血清能显著抑制A549细胞增殖(P<0.05),降低细胞克隆形成和迁移能力(P<0.05),阻滞细胞周期从G1期到S期转变,同时抑制Gli1、Ptch1、Smo和CyclinD1 mRNA和蛋白表达(P<0.05),且与Vismodegib有良好的协同作用.结论 理肺消积丸可显著抑制A549增殖和迁移能力,作用机制可能与其调控Hedgehog通路阻滞细胞周期有关.
Abstract
Objective To investigate the effect of Lifei-Xiaoji pills on A549 cells and the regulation of Hedgehog pathway.Methods MTT assay was used to select the optimal intervention concentration of Lifei-Xiaoji containing serum on A549 cells.Then A549 cells were divided into 6 groups:Control group,Lifei-Xiaoji containing serum group,Hedgehog pathway inhibitor group(Vismodegib group),Cisplatin group,Lifei-Xiaoji containing serum + Vismodegib group,Lifei-Xiaoji containing serum + cisplatin group.After different interventions,cell proliferation ability was detect by CCK-8 assay,cell cloning into pellets was observed by clonal formation assay,migration ability was observed by transwell assay,the change of cell cycle ratio was observed by flow cytometry,mRNA and protein expressions of Gli1,Ptch1,Smo and CyclinD1 in Hedgehog pathway were detected by qRT-PCR and Western blot,respectively.Results The serum containing Lifei-Xiaoji pills with 10%concentration could significantly inhibit the proliferation of A549 cells(P<0.05),decrease cell clonogenesis and migration ability(P<0.05),block the cell cycle transition from G1 phase to S phase,inhibit Gli1,Ptch1,Smo and CyclinD1 mRNA and protein expression(P<0.05),and has good synergistic effect with Vismodegib.Conclusion Lifei-Xiaoji Pills can significantly inhibit A549 cell proliferation and migration,and the mechanism may be related to the down-regulation of Hedgehog pathway and cell cycle arrest.
关键词
非小细胞肺癌/理肺消积丸/A549细胞/Hedgehog通路/细胞周期Key words
Non-small cell lung cancer/Lifei-Xiaoji pills/A549 cells/Hedgehog pathway/Cell cycle引用本文复制引用
基金项目
河南省卫生健康委国家中医临床研究基地科研专项(2021JDZY033)
河南省卫生健康委河南省中医药科学研究专项(2022ZY1022)
河南省自然科学基金青年项目(232300420272)
出版年
2024
国家科技期刊平台
NSTL
万方数据