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[摘要]
目的 研究片仔癀对肺癌A549细胞增殖、迁移、侵袭的抑制作用及其机制。方法 以人肺癌A549细胞作为体外实验对象,分别用不同质量浓度的片仔癀处理,MTT检测细胞增殖变化,Transwell小室测定细胞侵袭和迁移变化,Western blotting检测细胞中侵袭、迁移蛋白MMP-9、MMP-2和上皮细胞-间充质转化(EMT)蛋白E-cadherin、vimentin及Akt信号通路蛋白Akt磷酸化水平。用Akt信号通路激活剂和片仔癀处理肺癌细胞,检测激活Akt信号通路对片仔癀抗肺癌细胞增殖、侵袭和迁移的作用。结果 100 μg/mL的片仔癀对肺癌细胞增殖能力没有影响(P>0.05),200 μg/mL的片仔癀处理可以明显降低肺癌细胞增殖能力(P<0.05)。100、200 μg/mL的片仔癀处理后的肺癌细胞侵袭和迁移能力均降低,细胞中MMP-2和MMP-9蛋白表达水平下降,E-cadherin蛋白水平升高,vimentin蛋白水平降低,同时细胞中Akt磷酸化水平也下降(P<0.05)。Akt信号通路激活剂处理可以明显减弱片仔癀对肺癌细胞增殖、侵袭、迁移的抑制作用(P<0.05)。结论 片仔癀具有降低肺癌细胞增殖、迁移、侵袭能力,作用机制与抑制Akt信号通路的激活有关。
[Key word]
[Abstract]
Objective To study the inhibitory effect of Pien Tze Huang on proliferation, migration, and invasion of lung cancer cells and explore its mechanism. Methods Human lung cancer cell A549 was used as the experimental object in vitro, and the cells were treated with different concentrations of Pien Tze Huang. MTT was used to detect cell proliferation, and the changes of cell invasion and migration were measured by Transwell assay. Western blot was used to detect the phosphorylation of invasive migration proteins MMP-9, MMP-2 and EMT proteins E-cadherin, vimentin and Akt signaling pathway proteins. Lung cancer cells were treated with Akt signaling pathway activator and tablets to detect the effect of activating Akt signaling pathway on the proliferation, invasion, and migration of lung cancer cells. Results Pien Tze Huang (100 μg/mL tablets) had no effect on the proliferation of lung cancer cells, the proliferative ability of lung cancer cells could be significantly reduced by 200 μg/mL Pien Tze Huang. The invasion and migration ability of lung cancer cells treated with 100 and 200 μg/mL tablets were decreased, the protein expressions of MMP-2 and MMP-9 in cells were decreased, the level of E-cadherin protein in cells was increased, the level of vimentin protein was decreased, at the same time, the phosphorylation level of Akt was decreased. Akt signaling pathway activator treatment could significantly reduce the inhibitory effect of Pien Tze Huang on proliferation, invasion and migration of lung cancer cells (P<0.05). Conclusion Pien Tze Huang decreased the proliferation, migration and invasion of lung cancer cells, the mechanism is related to the inhibition of the activation of Akt signaling pathway.
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[基金项目]
国家自然科学基金资助项目(81473638);2017年度河南省医学科技攻关计划项目(201702257)