| 汪玉琪,来岳标,姚庆华.姜黄素对TGF-β诱导的A549细胞上皮间质转化的影响[J].浙江中西医结合杂志,2016,26(1): |
| 姜黄素对TGF-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 |
| 投稿时间：2015-06-30 修订日期：2015-12-02 |
| DOI： |
| 中文关键词: 姜黄素 上皮间质转化 Wnt/β-catenin信号通路 |
| 英文关键词:Curcumin Epithelial-Mesenchymal Transition Wnt/β-catenin signaling pathway |
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| 中文摘要: |
| 目的 研究姜黄素对转化生长因子-β（transforming growth factor-β,TGF-β)诱导的肺腺癌A549细胞的上皮间质转化（Epithelial-Mesenchymal Transition，EMT）的作用及可能的机制。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。实验分为4组：对照（Control）组、TGF-β组（TGF-β5ng/mL）、TGF-β+姜黄素组（TGF-β5ng/mL+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，采用免疫荧光和Western印迹法实验检测各组EMT相关标记物E-钙粘蛋白、N-钙粘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、4 μmol/L）作用后EMT表达情况。结果 MTT实验表明姜黄素对A549细胞的抑制作用呈时间和剂量浓度依赖性，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）；细胞运动迁移及侵袭浸润实验表明TGF-β能显著促进细胞的运动迁移和侵袭浸润，而姜黄素对其表现出抑制作用（P<0.05）。免疫荧光实验及免疫蛋白印迹实验表明TGF-β能下调E--钙粘蛋白的表达，上调波形蛋白及N--钙粘蛋白的表达，伴有P-GSK3β，β-catenin和c-Myc的表达增加以及Snail蛋白表达增加，而GSK3β表达无明显变化，而姜黄素表现出对这种现象的一个抑制作用。同时，XAV939对TGF-β诱导的EMT表现出抑制作用。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EMT。 |
| 英文摘要: |
| Objective To study the effect and mechanism of Curcumin on TGF-β-Induced EMT in A549 lung adenocarcinoma cells. Methods Human A549 lung adenocarcinoma cells was used to detect inhibition rate of curcumin by MTT assay. The study was divided into four groups: control group, TGF-β group(treated with TGF-β 5ng/mL), TGF-β+curcumin group(treated with TGF-β 5ng/mL and curcumin 10μmol/L), curcumin group(treated with curcumin 10μmol/L). Transwell and cell invasion assays was used to detect the movement and infiltration ability of each group. Immunofluorescence and Western blot analysis was used to detect the expression level of EMT related markers, such as E-cadherin, N-cadherin and Vimentin, also the expression level of P-GSK3β, GSK3β, β-catenin, c-Myc which is related to Wnt/β-catenin signaling pathway and Snail. In addition，the expression level of EMT effected by Wnt/β-catenin inhibitor XAV939 was tested. Results MTT assay showed that curcumin inhibited the proliferation of A549 cells along with the increase of the concentration and treatment time. When treated with 10μmol/L for 24h and 48h, the inhibition rate of cell proliferation was 12.21 ± 2.60% and 21.33 ± 3.25% (P<0.05), respectively; Transwell and cell invasion assays showed that TGF-β significantly stimulated cell invasion, an effect that was completely blocked by curcumin (P<0.05). In immunofluorescence and western blot experiments, TGF?β made a decreased expression of E-cadherin and an increased expression of vimentin and N-cadherin. Moreover, it promoted the expression of P-GSK3β, β-catenin, c-Myc proteins and snail, but made little changes in total of GSK3β. However, both of effects were blocked by curcumin. What’s more, XAV939 reversed TGF?β-induced EMT, expressed through the inhibition of N-cadherin, Vimentin expression and the increase of E-cadherin expression. Conclusion Curcumin inhibited EMT induced by TGF-β in A549 lung adenocarcinoma cells probably through inhibitting the Wnt/β-catenin signal. |
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