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

作　　者：刘素晓 张丛丛 孔德琪 黄慧敏 毛静[3] 王元元[1] 刘学芳[3] 冯素香[3] 李亚[1,3] LIU Suxiao;ZHANG Congcong;KONG Deqi;HUANG Huimin;MAO Jing;WANG Yuanyuan;LIU Xuefang;FENG Suxiang;LI Ya(The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Provincial Co-Construction and Collaborative Innovation Center of TCM Prevention and Treatment of Respiratory Diseases,Henan Key Laboratory of TCM Prevention and Treatment of Respiratory Diseases,Henan University of Chinese Medicine,Zhengzhou 450046,China;The First Clinical Medical College of Henan University of Traditional Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [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

出　　处：《世界科学技术-中医药现代化》2024年第2期320-327,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：河南省卫生健康委员会国家中医临床研究基地科研专项(2021JDZY033):理肺消积丸经Hedgehog/GLI通路调控细胞增殖和凋亡抑制肺癌小鼠肿瘤生长的机制研究,负责人:刘素晓;河南省卫生健康委员会河南省中医药科学研究专项(2022ZY1022):理肺消积丸经lnc RNA HOTAIR/Wnt/β-catenin轴调控EMT抗肺癌的体外活性机制研究,负责人:刘素晓;河南省科学技术厅河南省自然科学基金青年项目(232300420272):理肺消积丸通过USP47/BACH1轴调控Warburg效应抑制肺腺癌细胞增殖和迁移的机制研究,负责人:刘素晓。

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。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 

分 类 号：R285.5[医药卫生—中药学]