益肺解毒汤对人肺腺癌A549细胞侵袭迁移的影响及相关机制  

Intervention Effect and Mechanism of Yifei Jiedu Decoction in Invasion and Metastasis of Lung Cancer Cell A549

作　　者：王珊珊[1] 赵新鲜 郭立中[1] WANG Shan-shan ZHAO Xin-xian GUO Li-zhong(Nanjing University of Chinese Medicine, Nanjing 210029, China The Affiliated Jiangning Hospital of Nanjing Medical University, Nanjing 210029, China)

机构地区：[1]南京中医药大学,南京210029 [2]南京市江宁医院,南京210029

出　　处：《中国实验方剂学杂志》2017年第22期108-113,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江苏省普通高校科研创新计划项目(KYZZ16_0408)

摘　　要：目的:观察益肺解毒汤对肺癌A549细胞侵袭转移的影响,并探讨该方对细胞上皮-间充质转化(EMT)的影响及其作用机制。方法:常规培养肺癌A549细胞,Co Cl2化学模拟肺癌A549细胞缺氧微环境,设空白组,以150μmol·L^(-1)Co Cl2,150μmol·L^(-1)Co Cl2+15 g·L^(-1)益肺解毒汤,15 g·L^(-1)益肺解毒汤处理细胞,镜下观察细胞形态学改变,transwell实验观察细胞侵袭能力;细胞划痕实验测细胞迁移能力;蛋白免疫印迹法(Western blot)测定A549细胞上皮细胞-间充质转化(EMT)过程中上皮细胞、间质细胞标志物E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)表达,转录因子Snail蛋白表达,信号分子β-链蛋白(β-catenin)、磷酸化糖原合成酶激酶-3(p-gsk-3β)表达。结果:缺氧可诱导A549细胞发生形态改变,益肺解毒汤可抑制缺氧诱导的形态改变。与空白组比较,Co Cl2组细胞侵袭和迁移能力增强(P<0.05);与Co Cl2组比较,Co Cl2+益肺解毒汤组的细胞侵袭能力明显减弱(P<0.01),迁移能力亦减弱(P<0.05)。与空白组比较,Co Cl2组细胞EMT相关蛋白,E-cadherin表达下调,Vimentin,Snail表达上调(P<0.05),说明缺氧使细胞发生了EMT;与Co Cl2组比较,Co Cl2+益肺解毒汤组可上调Ecadherin,p-gsk-3β,下调Vimentin,Snail,β-catenin蛋白表达(P<0.05)。结论:益肺解毒汤能抑制缺氧诱导的A549细胞的形态变化、侵袭及迁移能力,还能抑制缺氧诱导的EMT的发生,其机制可能与Wnt/β-catenin通路有关。Objective： To observe the intervention effect and mechanism of Yifei Jiedu decoction on the invasion and metastasis of lung cancer adenocarcinoma A549 cells. Method： A549 cells were cultured and treated with 150 μmol·L^-1Co Cl2,150 μmol·L^-1Co Cl2＋ 15 g·L^-1Yifei Jiedu decoction,and 15 g·L^-1Yifei Jiedu decoction,respectively. Untreated cells were used for normal control. Morphological changes were observed under inverted microscope. Wound scratch assay were performed to further identify if Yifei Jiedu decoction could reduce the migration ability of A549 cells. Transwell was used to explore the intervention effect of Yifei Jiedu decoction on the invasion and movement of A549 cells. Expressions of E-cadherin,Vimentin,Snail,β-catenin and p-gsk-3βwere examined by Western blot. Result： Co Cl2 can induce morphological changes in A549 cells,which were accompanied by up-regulation of E-cadherin expression and down-regulation of Vimentin. Wound scratch assay indicated that Yifei Jiedu decoction could reduce the migration ability of A549 cells. Transwell assay showed that Yifei Jiedu decoction could inhibit the invasion of A549 cells（ P〈0. 05）. Western blot showed that Yifei Jiedu decoction could up-regulate the expressions of E-cadherin,p-gsk-3β protein and down-regulate the expressions of Vimentin,Snail and β-catenin protein respectively（ P〈0. 05）. Conclusion： Yifei Jiedu decoction could inhibit the migration and invasion of human adenocarinoma A549 cells via regulation of β-catenin expression.

关 键 词：中药复方 益肺解毒汤 上皮-间充质转化 Wnt/β-链蛋白信号通路 

分 类 号：R285.5[医药卫生—中药学]