缺氧条件下肺岩宁方抑制TC2N的表达对肺癌细胞培养基诱导的HUVEC细胞增殖、迁移侵袭和管腔形成的影响  

Influences of Feiyanning Formula Inhibiting the Expression of TC2N on the Proliferation,Migration,Invasion and Lumen Formation of HUVEC Cells Induced by Lung Cancer Cell Culture Medium Under Hypoxia

作　　者：王薇 徐振晔[1] 邓海滨[1] 马玥[1] 王中奇[1] 王立芳[1] Wang Wei;Xu Zhenye;Deng Haibin;Ma Yue;Wang Zhongqi;Wang Lifang(Longhua Hospital Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032

出　　处：《世界科学技术-中医药现代化》2023年第4期1250-1258,共9页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：上海市教育委员会科研创新计划重大项目(2017-01-07-00-10-E00064):源于精气理论的肺岩宁方抗肺癌生长与转移作用机制的研究,负责人:邓海滨

摘　　要：目的探讨缺氧条件下肺岩宁方(FYN)对肺癌细胞培养基诱导的人脐静脉内皮细胞(HUVEC)增殖、迁移侵袭和管腔形成的影响。方法使用不同浓度的肺岩宁方(200-6400μg·mL^(-1))处理HUVEC细胞,CCK-8法检测细胞存活率,计算其半数抑制浓度(IC_(50)),确定最佳干预浓度。使用A549细胞培养基在缺氧条件下诱导血管新生模型,将HUVEC细胞分为对照组(Control)、模型组(Model)、800μg·mL^(-1)肺岩宁方组(FYN)、100μg·mL^(-1)贝伐珠单抗组(Bevacizumab,阳性对照)、800μg·mL^(-1)肺岩宁方+慢病毒载体组(FYN+vector)、800μg·mL^(-1)肺岩宁方+TC2N过表达组(FYN+TC2N),每组设置6个复孔。分别检测细胞增殖、迁移与侵袭,管腔生成实验评价血管形成,qRT-qPCR检测HIF-1α、TC2N基因水平,Western blot法检测HIF-1α、TC2N、E-cadherin、Vimentin、Snail蛋白表达。结果肺岩宁方对HUVEC细胞的IC_(50)为893.00μg·mL^(-1);与对照组比较,模型组细胞存活率、细胞迁移与侵袭能力、管腔形成节点数、Vimentin、Snail、HIF-1α、TC2N基因及蛋白表达升高,E-cadherin表达降低(P<0.05);与模型组比较,肺岩宁方组与贝伐珠单抗组细胞存活率、细胞迁移与侵袭能力、管腔形成节点数、Vimentin、Snail、HIF-1α、TC2N基因及蛋白表达降低,E-cadherin表达升高(P<0.05);与肺岩宁方+慢病毒载体组比较,肺岩宁方+TC2N过表达组细胞存活率、细胞迁移与侵袭能力、管腔形成节点数、Vimentin、Snail、TC2N基因及蛋白表达升高,E-cadherin表达降低(P<0.05)。结论肺岩宁方可通过抑制TC2N的表达,抑制HUVEC细胞增殖、迁移、侵袭与血管生成。Objective To investigate the influences of Feiyanning formula(FYN)on the proliferation,migration,invasion and lumen formation of human umbilical vein endothelial cells(HUVEC)induced by lung cancer cell culture medium under hypoxia.Methods HUVEC cells were treated with different concentrations of Feiyanning formula(200-6400μg·mL^(-1)),the cell viability was detected by CCK-8 method,and the median inhibitory concentration(IC_(50))was calculated to determine the optimal intervention concentration.A549 cell culture medium was used to induce angiogenesis model under hypoxia,and HUVEC cells were separated into Control group,Model group,800μg·mL^(-1)Feiyanning formula group(FYN),and 100μg·mL^(-1)bevacizumab group(positive control),800μg·mL^(-1)Feiyanning formula+lentiviral vector group(FYN+vector),800μg·mL^(-1)Feiyanning formula+TC2N overexpression group(FYN+TC2N),each group included 6 duplicate wells.Cell proliferation,migration and invasion were detected respectively,Lumenogenesis assays were applied to evaluate vessel formation,qRT-qPCR was applied to detect the levels of HIF-1α,TC2N gene,Western blot was applied to detect the protein expression of HIF-1α,TC2N,E-cadherin,Vimentin,and Snail.Results The IC_(50)of Feiyanning formula on HUVEC cells was 893.00μg·mL^(-1);compared with the control group,the cell survival rate,cell migration and invasion ability,the number of lumen formation nodes,the expression levels of Vimentin,Snail,HIF-1α,TC2N gene and protein in the model group were obviously increased,the expression level of E-cadherin was obviously decreased(P<0.05);compared with the model group,the cell survival rate,cell migration and invasion ability,the number of lumen formation nodes,the expression levels of Vimenti,Snail,HIF-1α,TC2N gene and protein in the Feiyanning formula group and bevacizumab group were obviously decreased,the expression level of Ecadherin was obviously increased(P<0.05);compared with Feiyanning formula+lentiviral vector group,the cell survival rate,cell migration and invasi

关 键 词：肺岩宁方 串联C2结构域核蛋白 人脐静脉内皮细胞 增殖 迁移 管腔形成 

分 类 号：R285.5[医药卫生—中药学]