姜黄素对转化生长因子-β诱导的A549细胞上皮间质转化的影响  

Effect of Curcumin on Epithelial-Mesenchymal Transition Induced by TGF-β in A549 lung adenocarcinoma cells

作　　者：汪玉琪[1] 来岳标[1] 姚庆华[2] 

机构地区：[1]浙江中医药大学第一临床医学院,杭州310053 [2]浙江省肿瘤医院中西医结合科,杭州310022

出　　处：《浙江中西医结合杂志》2016年第1期22-26,99,100,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的研究姜黄素对转化生长因子-β(TGF-β)诱导的肺腺癌A549细胞上皮间质转化(EMT)作用及可能机制。方法选取体外培养肺腺癌A549细胞采用MTT法检测姜黄素细胞抑制率。实验分对照组、TGF-β组(TGF-β5ng/m L)、TGF-β+姜黄素组(TGF-β5ng/m L+姜黄素10μmol/L)、姜黄素组(姜黄素10μmol/L)。采用Transwell细胞迁移及浸润实验检测各组的运动迁移和侵袭浸润的能力,免疫荧光和Western印迹法实验检测各组EMT相关标记物E-钙黏蛋白、N-钙黏蛋白和波形蛋白的表达水平与Wnt/β-catenin信号通路相关P-GSK3β、GSK3β、β-catenin、c-Myc蛋白以及Snail蛋白表达水平,同时检测Wnt/β-catenin抑制剂XAV939(1、2、4μmol/L)作用后EMT表达情况。结果 MTT实验表明,10μmol/L姜黄素作用A549细胞24h和48h时,细胞增殖抑制率分别为(12.21±2.60)%和(21.33±3.25)%(P<0.05);TGF-β能显著促进细胞的运动迁移和侵袭浸润,下调E-钙黏蛋白的表达,上调波形蛋白及N-钙黏蛋白的表达,伴有P-GSK3β、β-catenin和c-Myc的表达增加以及Snail蛋白表达增加,GSK3β表达无明显变化,而姜黄素可以抑制TGF-β的作用(P<0.05)。同时,XAV939对TGF-β诱导的EMT也有抑制作用。结论姜黄素可能通过抑制Wnt/β-catenin信号通路来抑制TGF-β诱导的A549细胞上皮间质转化。Objective To investigate the effect of curcumin on transforming growth factor β（TGF-β）-inducedepithelial-mesenchymal Transition（EMT） in A549 lung adenocarcinoma cells and the underlying mechanism. Methods Human A549 lung adenocarcinoma cells was used to detect inhibition rate of curcumin by MTT assay. The studywas divided into 4 groups： control group, TGF-β group（treated with TGF-β5ng/m L）, TGF-β＋curcumin group（treated with TGF-β 5ng/m L and curcumin 10μmol/L）, curcumingroup（treated with curcumin 10μmol/L）.Transwell and cell invasion assays were used to detect the movement and infiltration ability of each group. Immunofluorescence and Western blot analysis were used to detect the expression level of EMT related markers such as E-cadherin, N-cadherin and vimentin and the expression level of Wnt/β-cateninsignaling pathway-related P-GSK3β,GSK3β, β-catenin and c-Mycand Snail protein. The expression level of EMT effected by Wnt/β-catenin inhibitor XAV939 was also determined. Results MTT assay showed that 10μmol/Lcurcumin inhibited the proliferation of A549 cells along with the increase of the treatment time（24h： 12.21%±2.60%; 48h： 21.33%±3.25%; P〈0.05）.TGF-β significantly stimulated cell movement and invasion, reduced the expression of E-cadherin, increased the expression of vimentin and N-cadherin, promoted the expression of P-GSK3β, β-catenin, c-Myc proteins and Snail, but made little changes in total of GSK3β. These effectsof TGF-β on A549 cells werecompletely blocked by curcumin（P〈0.05）. XAV939 reversed TGF-β-induced EMT. Conclusion Curcumin inhibited EMT induced by TGF-β in A549 lung adenocarcinoma cells probably through inhibiting the Wnt/β-catenin signaling pathway.

关 键 词：肺腺癌A549细胞 上皮间质转化 姜黄素 WNT/Β-CATENIN信号通路 

分 类 号：R285[医药卫生—中药学]