| 阿托伐他汀对转化生长因子β1诱导的肺癌A549细胞上皮间质转化过程的影响 |
| 投稿时间：2017-02-13 修订日期：2017-04-20 点此下载全文 |
| 引用本文：刘洋.阿托伐他汀对转化生长因子β1诱导的肺癌A549细胞上皮间质转化过程的影响[J].医学研究杂志,2017,46(12):41-46 |
| DOI：
10.11969/j.issn.1673-548X.2017.12.011 |
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| 中文摘要:目的 研究阿托伐他汀对TGF-β1诱导的A549细胞上皮间质转化（EMT）过程的影响。方法 体外培养肺癌A549细胞，阿托伐他汀以不同的浓度（0.5、1、2μmol/L）预处理A549细胞24h，随后，在无血清条件下加入TGF-β1（1ng/ml），共同作用48h，Western blot法检测EMT过程标志性蛋白E-钙黏蛋白（E-cadherin）、间质细胞标志蛋白波形蛋白（vimentin）的表达。体外培养A549细胞，阿托伐他汀预处理24h后，将A549细胞种板于Transwell迁移小室，观察阿托伐他汀对TGF-β1诱导的EMT过程的细胞迁移能力的影响。异硫氰酸荧光素标记的鬼笔环肽用来显影肌动蛋白纤维，共聚焦荧光显微镜观察肌动蛋白应力纤维的变化，研究阿托伐他汀对TGF-β1诱导的肌动蛋白应力纤维的影响。Western blot法检测EMT过程转录因子的表达变化。结果 阿托伐他汀预处理的A549细胞在TGF-β1的作用下向间质样细胞形态的转变过程受到抑制。以TGF-β1诱导EMT过程，阿托伐他汀处理组细胞E-Cadherin的表达高于阳性对照组细胞，相反，阿托伐他汀处理组细胞vimentin的表达低于阳性对照组细胞，提示阿托伐他汀抑制A549细胞EMT过程。A549细胞细胞经阿托伐他汀预处理24h后，加入TGF-β1作用24h，阿托伐他汀处理组的细胞内应力纤维的数量低于阳性对照组细胞。Transwell迁移实验，TGF-β1刺激和不刺激细胞时，阿托伐他汀预处理24h的A549细胞迁移至下室的细胞数皆少于阴性对照组细胞。阿托伐他汀预处理的A549细胞在TGF-β1的作用下，转录因子ZEB1的表达低于阴性对照组细胞，且两组细胞的Snail和Slug的表达差异无统计学意义，而在TGF-β1的作用下Snail和Slug表达都逐渐增高。结论 阿托伐他汀能部分抑制TGF-β1诱导的A549细胞的上皮间质转化过程，使转录因子ZEB1的表达降低，肌动蛋白应力纤维束形成减少，TGF-β1诱导的细胞迁移受阻。 |
| 中文关键词:阿托伐他汀 非小细胞肺癌 上皮间质转化 TGF-β1 |
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| Atorvastatin Partially Inhibit the EMT Process Induced by TGF-β1 |
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| Abstract:Objective To investigate the effect of atorvastatin on the EMT process of A549 cells induced by TGF-β1.Methods A549 cells were cultured in vitro and exposed at different concentration in atorvastatin (0.5,1,2μmol/L). 24h later, the TGF-β1 was added to the supernatants with the concentration of 1 ng/ml. After another 48 hours co-exposed to atorvastatin and TGF-β1, the levels of EMT marker proteins (E-cadherin,vimentin) were evaluated by Western blot. Atrovastatin pretreated A549 cells were transported to transwell upper chambers. Cells on the lower surface of the filters were examined to evaluated effect of atorvastatin on the ability of migration induced by TGF-β1. FITC-phalloidine was used to make actin fibers visible by confocal laser scanning microscope and to determine the effect of atorvastasin on actin fibers induced by TGF-β1.The transcriptional factors inhibiting the transcription of E-cadherin were evaluated by Western blot.Results There was no effect of atorvastatin alone on the expression of E-cadherin and vimentin evaluated by western blot. The morphological transiton to fibroblast-like cell morphology of A549 cells pretreated with atorvastatin was inhibited under the effect of TGF-β1. During the EMT process induced by TGF-β1,compared with positive control cells (exposed to TGF-β1 only),A549 cells pretreated with atorvastatin had a higher expression of E-cadherin, which was the marker protein of epithelial cells. On the contrary, the expression of vimentin of A549 cells pretreated with atorvastatin was lower. This result indicated that atorvastatin inhibit the EMT process induced by TGF-β1. After exposing to TGF-β1 for 24 hours, compared with positive control cells, the actin stress fibers of cells (A549 cells and NCI-H1975 cells) pretreated with atorvastatin for 24 hours were more less. Transwell experiment indicated that compared with negative control, atorvastatin could reduce the number of cells migrating to the lower surface of filters. When exposing to TGF-β1, compared with positive control cells, atorvastatin could reduce the number of cells migrating to the lower surface of filters induced by TGF-β1 either. When exposing to TGF-β1, compared with A549 cells without atorvastatin pretreatment, the expression of ZEB1 of A549 cells pretreated by atorvastatin was lower. The expression of snail and slug were similar between two A549 cells groups. In both groups snail and slug were upregulated at different time spot by TGF-β1 stimulation.Conclusion Atorvastatin can partially inhibit the EMT process of A549 Cells induced by TGF-β1, reduce the expression of transcription factor ZEB1. The generation of actin stress fibers is weakened. The migration ability increased by TGF-β1 is inhibited by atorvastatin. |
| keywords:Atorvastatin Non-small cell lung cancer (NSCLC) Epithelial-mesenchymal transition TGF-β1 |
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