紫菀水提物对人肺癌A549细胞增殖、侵袭及裸鼠成瘤能力的影响及作用机制探讨  

Effects and aster water extract on proliferation and invasion of human lung cancer A549 cells,and tumorigenesis ability of nude mice

作　　者：姚平波 刘雨露 朱子贵 赵红[3] 张建新[4] YAO Ping-bo;LIU Yu-lu;ZHU Zi-gui;ZHAO Hong;ZHANG Jian-xin(Changsha Civil Affairs Vocational and Technical College,Changsha 410004,China;Department of Thoracic Surgery,Nanhua Hospital Affiliated to Nanhua University,Hengyang 421002,China;Department of Basic Nursing,School of Nursing,Nanhua University,Hengyang 421001,China;Department of Critical Care Medicine,Nanhua Hospital Affiliated to Nanhua University,Hengyang 421002,China)

机构地区：[1]长沙民政职业技术学院,长沙410004 [2]南华大学附属南华医院重症医学科,衡阳421002 [3]南华大学护理学院基础护理教研室,衡阳421001 [4]南华大学附属南华医院胸外科,衡阳421002

出　　处：《药物分析杂志》2022年第3期380-386,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：湖南省自然科学基金项目(2018JJ6122)。

摘　　要：目的:探讨紫菀水提物对人肺癌A549细胞增殖、侵袭及裸鼠成瘤能力的影响及作用机制。方法:将体外培养MRC-5和A549细胞分别与不同浓度AWE(0、0.6、1.25、2.5、5、10、20、40、80、160、320 g·L^(-1))孵育,根据MTT实验设置A549细胞为AWE 0 g·L^(-1)组、AWE 10·L^(-1)组、AWE 20·L^(-1)组和AWE 40·L^(-1)组。CCK-8法检测各组细胞增殖,Transwell法检测侵袭细胞数。采用Wnt/β-catenin信号通路抑制剂XAV939处理A549细胞,Western blot检测各组细胞Wnt-1、β-catenin、血管内皮生长因子(VEGF)和Ki-67蛋白表达。皮下种植A549细胞悬液(5×10^(6)个·mL^(-1))0.2 mL,经AWE(灌胃5 g·kg^(-1)·d^(-1),4周)干预后,进行移植瘤组织称重,免疫组化检测VEGF和Ki-67表达。结果:AWE对MRC-5细胞无明显抑制作用(P>0.05),对A549细胞的抑制作用呈药物浓度依赖性,IC_(50)=24.81 g·L^(-1),随着AWE浓度增加,AWE浓度≥20 g·L^(-1)时,细胞存活率下降(P<0.05)。AWE 20 g·L^(-1)组和AWE 40 g·L^(-1)组A549细胞增殖活性和侵袭细胞数低于AWE 0 g·L^(-1)组(P<0.05)。AWE 40 g·L^(-1)组和XAV939组A549细胞Wnt-1、β-catenin、Ki67和VEGF蛋白表达低于对照组(P<0.05);AWE 40+XAV939组A549细胞Wnt-1、β-catenin、Ki67和VEGF蛋白表达低于AWE 40 g·L^(-1)组和XAV939组(P<0.05)。裸鼠成瘤实验结果显示,AWE组裸鼠移植瘤质量低于对照组(P<0.05),且移植瘤组织中Ki67和VEGF蛋白表达低于对照组(P<0.05)。结论:AWE可以抑制肺癌细胞增殖和侵袭,抑制移植瘤生长,可能与抑制Wnt/β-catenin信号通路有关。Objective:To explore the effects and mechanisms of aster water extract(AWE)on the proliferationand invasion of human lung cancer A549 cells,and the tumorigenesis capability of nude mice.Methods:MRC-5 and A549 cells were cultured in vitro and incubated with different concentrations of AWE(0,0.6,1.25,2.5,5,10,20,40,80,160,320 g·L^(-1)).In the MTT assay,A549 cells were divided into AWE 0 g·L^(-1) group,AWE 10 g·L^(-1) group,AWE 20 g·L^(-1) group and AWE 40 g·L^(-1) group,and the cell proliferation rate in each group was detected by CCK-8.The number of invasion cells was detected using Transwell method.A549 cells were treated with Wnt/β-catenin signaling pathway inhibitor XAV939.The expression ofβ-catenin,vascular endothelial growth factor(VEGF)and Ki-67 proteins in each group was detected by Western blot.About 0.2 mL of A549 cells suspension(5×10^(6) cell·mL^(-1))was subcutaneously implanted.After the intervention of AWE(5 g·kg^(-1)·d^(-1) intragastric administration,4 weeks),transplanted tumor tissues was weighed.The expression of VEGF and Ki-67 was detected by immunohistochemistry.Results:There was no significant inhibitory effect of AWE on MRC-5 cells(P>0.05),the inhibitory effect on A549 cells showed concentration-dependence(IC_(50)=24.81 g·L^(-1)).With the increase of AWE concentration,when AWE concentration was not lower than 20 g·L^(-1),survival rate of cells was decreased(P<0.05).The proliferation activity of A549 cells and number of invasion cells in AWE 20 g·L^(-1) group and AWE 40 g·L^(-1) group were lower than those in AWE 0 g·L^(-1) group(P<0.05).The expressions of Wnt-1,β-catenin,Ki67 and VEGF proteins in A549 cells of AWE 40 g·L^(-1) group and XAV939 group were lower than those in control group(P<0.05),which were lower in AWE 40 g·L^(-1)+XAV939 group than AWE 40 g·L^(-1) group and XAV939 group(P<0.05).The results of tumorigenicity assay in nude mouse showed that the mass of transplanted tumors in AWE group was lower than that in control group(P<0.05),and the expression of Ki67 and

关 键 词：紫菀水提物 肺癌 增殖 侵袭 WNT/Β-CATENIN信号通路 

分 类 号：R917[医药卫生—药物分析学]