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柠檬酚对人肝癌HepG2细胞增殖、迁移和骨架相关蛋白表达的影响及其与蛋白作用方式研究
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| 篇名： | 柠檬酚对人肝癌HepG2细胞增殖、迁移和骨架相关蛋白表达的影响及其与蛋白作用方式研究 |
| TITLE： | Study on the Effects of Citrusinol on the Proliferation ，Migration and the Expression of Skeleton-related Proteins of Human Hepatocellular Cells HepG 2 and Its Interaction Mode with Skeleton-related Proteins |
| 摘要： | 目的：研究柠檬酚对人肝癌HepG2细胞增殖、迁移及骨架相关蛋白表达的影响，并探讨其与骨架相关蛋白的相互作用方式。方法：采用CCK-8法检测不同浓度柠檬酚（12.5、25、50、100、200μmol/L）作用24h后对HepG2细胞增殖的影响。将HepG2细胞分为阴性对照组和柠檬酚低、高浓度组（50、100μmol/L柠檬酚），加入相应药物作用24h后，采用划痕试验检测细胞的迁移能力并计算细胞迁移率，采用逆转录-聚合酶链式反应和Westernblotting法检测细胞中聚合纤维状肌动蛋白（F-actin）、β-微管蛋白（β-tubulin）及埃兹蛋白（Ezrin）的mRNA和蛋白表达水平。利用分子对接软件Schrodinger2015分析柠檬酚与上述3种蛋白的分子作用方式。结果：柠檬酚对HepG2细胞增殖具有显著抑制作用（P＜0.05或P＜0.01），且呈剂量依赖趋势。与阴性对照组比较，柠檬酚低、高浓度组细胞迁移率和F-actin、β-tubulin、Ezrin的mRNA及蛋白表达水平显著降低（P＜0.05或P＜0.01）。分子对接结果显示，柠檬酚可与上述3种细胞骨架蛋白形成氢键和疏水键。结论：柠檬酚可抑制肝癌HepG2细胞的增殖和迁移；其作用机制可能与下调F-actin、β-tubulin、Ezrin的mRNA及蛋白表达有关；其与骨架相关蛋白的作用方式可能是形成氢键或疏水键。 |
| ABSTRACT： | OBJECTIVE：To study the effects of citrusinol on proliferation ，migration and the expression of skeleton-related proteins of human hepatocellular cells HepG 2，and to investigate its interaction mode with skeleton-related proteins. METHODS ： CCK-8 assay was used to detect the effects of different concentrations （12.5，25，50，100，200 μmol/L）of citrusinol on the proliferation of HepG 2 cells for 24 h. HepG 2 cells were divided into negative control group ，citrusinol low-concentration and high-concentration groups （50，100 μmol/L citrusinol）. After treated for 24 h，the migration of HepG 2 cells was detected by cell scratch test ；cell migration rate was calculated. mRNA and protein expression of F-actin ，β-tubulin and Ezrin in HepG 2 cells were determined by RT-PCR and Western blotting assay. Molecular docking software Schrodinger 2015 was used to analyze the interaction mode between citrusinol and above 3 kinds of proteins. RESULTS ：Citrusinol showed significant inhibition effect on the proliferation of HepG 2 cells （P＜0.05 or P＜0.01），in dose-dependent trend. Compared with negative control group ，cell migration， mRNA and protein expression levels of F-actin ， β-tubulin， Ezrin were decreased significantly in citrusinol low-concentration and high-concentration groups （P＜0.05 or P＜0.01）. Molecular docking results showed that the citrusinol could form hydrogen bond and hydrophobic bond with the above 3 skeleton-related proteins. CONCLUSIONS ：Citrusinol can inhibit the proliferation and migration of HepG 2 cells，the mechanism may be associated with the down-regulation of mRNA and protein expression of F-actin ，β-tubulin and Ezrin. The mode of its interaction with skeleton-related proteins may be the formation of hydrogen bond or hydrophobic bond. |
| 期刊： | 2020年第31卷第15期 |
| 作者： | 黄周锋，胡筱希，陆国寿，黄建猷，谭晓 |
| AUTHORS： | HUANG Zhoufeng ，HU Xiaoxi ，LU Guoshou ，HUANG Jianyou ，TAN Xiao |
| 关键字： | 柠檬酚；人肝癌HepG2细胞；增殖；迁移；骨架相关蛋白；分子对接 |
| KEYWORDS： | Citrusinol；HepG2 cells；Proliferation；Migration；Skeleton-related proteins ；Molecule docking |
| 阅读数： | 294 次 |
| 本月下载数： | 7 次 |
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