摘要
目的:探讨刺槐素对非小细胞肺癌(non-small cell lung cancer,NSCLC)A549细胞迁移、侵袭和血管生成的影响及其分子机制。方法:MTT实验检测不同浓度刺槐素对A549细胞活力的影响并计算刺槐素IC 50;采用不同浓度刺槐素处理A549细胞,蛋白免疫印迹分别检测血管内皮生长因子(vascular endothelial growth factor,VEGF)、E-钙黏蛋白(E-cadherin)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)相对表达水平,划痕实验、Transwell侵袭实验、免疫荧光和小管生成实验分别检测细胞迁移、侵袭及血管生成能力;A549细胞转染过表达或敲减VEGF质粒,蛋白免疫印迹检测分别检测VEGF、E-cadherin和MMP-9相对表达水平,并进一步检测细胞迁移、侵袭及血管生成能力。结果:与0μmol/L组相比,10、20、30、40、50μmol/L刺槐素组A549细胞活力均明显降低(P均<0.05),且呈一定的浓度依赖性,IC 50为34.49μmol/L;与0μmol/L组相比,10、20、40μmol/L组A549细胞迁移、侵袭及血管生成能力明显减弱(P均<0.05),VEGF和MMP-9蛋白表达水平明显降低(P均<0.01),E-cadherin蛋白表达水平明显增加(P均<0.01);与空白对照或过表达阴性对照组相比,过表达组A549细胞MMP-9蛋白表达水平明显增加,E-cadherin蛋白表达水平明显降低,细胞迁移、侵袭及血管生成能力明显增强(P均<0.05);与空白对照或敲减阴性对照组相比,敲减组A549细胞MMP-9蛋白表达水平明显降低,E-cadherin蛋白表达水平明显增加,细胞迁移、侵袭及血管生成能力明显减弱(P均<0.05)。结论:刺槐素可能通过降低VEGF表达进而抑制NSCLC A549细胞迁移、侵袭及血管生成。
Objective:To investigate the effect and molecular mechanisms of acacetin on migration,invasion and angiogenesis of non-small cell lung cancer(NSCLC)A549 cells.Methods:The effects of various concentrations of acacetin on A549 cell viability were detected by MTT assay and IC 50 was calculated.The A549 cells were treated with various concentrations of acacetin,and the relative expression levels of vascular endothelial growth factor(VEGF),E-cadherin and matrix metalloproteinase-9(MMP-9)were detected by Western blotting.And the ability of cell migration,invasion and angiogenesis was detected by scratch wound healing,Transwell,immunofluorescence and tubulogenesis tests,respectively.VEGF plasmid was overexpressed or knocked down in A549 cells,and the relative expression levels of VEGF,E-cadherin and MMP-9 were detected by Western blotting,and the ability of the migration,invasion and angiogenesis of the cells were further evaluated.Results:Compared with 0μmol/L group,A549 cell viability in 10,20,30,40 and 50μmol/L acacetin groups was significantly decreased(P<0.01),and in a certain concentration-dependent manner,IC 50 was 34.49μmol/L.Compared with 0μmol/L group,the migration,invasion and angiogenesis of A549 cells in 10,20 and 40μmol/L acacetin groups were significantly decreased(all P<0.05),and the expression levels of VEGF and MMP-9 protein were significantly decreased(all P<0.01),while the expression level of E-cadherin protein was significantly increased(P<0.01).Compared with blank control or overexpression negative control group,the expression level of MMP-9 protein in A549 cells in overexpression group was significantly increased,while the expression level of E-cadherin protein was significantly decreased,and the ability of cell migration,invasion and angiogenesis was significantly enhanced(all P<0.05).Compared with blank control or knockdown negative control group,the expression level of MMP-9 protein in A549 cells in knockdown group was significantly decreased,while the expression level of E-cadherin protei
作者
闫庆韬
万斯傲
刘意
陆健
吴占敖
YAN Qingtao;WAN Siao;LIU Yi;LU Jian;WU Zhan′ao(Institute of Life Science,Jiangsu University,Zhenjiang Jiangsu 212013;School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013;Department of Stomatology,Jinling Hospital,Medical School of Nanjing University,Nanjing Jiangsu 210093,China)
出处
《江苏大学学报（医学版）》
CAS
2023年第6期461-469,共9页
Journal of Jiangsu University：Medicine Edition
基金
军区医学科技创新重点项目(417411,ZX3)
金山英才人才项目(2018)。
