地高辛通过下调AEG-1表达抑制人胃癌MKN45细胞迁移和侵袭

发布时间：2018-02-28 16:18

本文关键词： 地高辛星形细胞上调基因- 迁移侵袭胃癌 MKN细胞　出处：《生理学报》2017年02期 　论文类型：期刊论文

【摘要】：本研究旨在观察地高辛对人胃癌MKN45细胞迁移和侵袭能力的影响,并探讨其分子机制。取人胃癌MKN45细胞作为研究对象,采用Transwell法检测细胞迁移和侵袭能力,通过脂质体转染法将星形细胞上调基因-1(astrocyte elevated gene-1,AEG-1)sh RNA干扰质粒转染MKN45细胞以构建低表达AEG-1的细胞株,利用Western blot法检测基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)、E-钙粘蛋白(E-cadherin)和AEG-1等蛋白的表达变化。结果显示,地高辛可使MKN45细胞迁移率和侵袭率均显著下降(P0.05),下调MMP-9和AEG-1蛋白表达水平(P0.05)以及上调E-cadherin蛋白表达水平(P0.05),上述作用均具有剂量依赖性。经sh RNA干扰AEG-1基因表达后,MKN45细胞中AEG-1蛋白表达水平显著下降(P0.05);同时AEG-1干扰组MKN45细胞中E-cadherin蛋白表达水平显著升高(P0.05),细胞迁移率、侵袭率和MMP-9蛋白表达水平均显著下降(P0.05)。上述结果提示,地高辛可在体外剂量依赖性地抑制人胃癌MKN45细胞迁移和侵袭,这可能与地高辛抑制AEG-1蛋白表达,继而下调MMP-9蛋白表达和上调E-cadherin蛋白表达有关。
[Abstract]:The purpose of this study was to observe the effect of digoxin on the migration and invasion of human gastric cancer MKN45 cells, and to explore its molecular mechanism. Human gastric cancer MKN45 cells were used as research objects, and the migration and invasion ability of human gastric cancer MKN45 cells was detected by Transwell assay. The up-regulation gene of astrocyte elevated gene-1 and AEG-1sh RNA interference plasmid were transfected into MKN45 cells by liposome transfection to construct a cell line with low expression of AEG-1. The expression of matrix metalloproteinase-9 (MMP-9) and E-cadherin (E-cadherin) and AEG-1 were detected by Western blot. Digoxin significantly decreased the migration and invasion of MKN45 cells, down-regulated the expression level of MMP-9 and AEG-1 protein (P0.05) and up-regulated the expression level of E-cadherin protein in a dose-dependent manner. These effects were in a dose-dependent manner. The expression of AEG-1 gene was interfered by sh RNA. The expression of E-cadherin in MKN45 cells with AEG-1 interference was significantly higher than that in P0.05 cells, and the cell migration rate was significantly increased in AEG-1 interference group, and the expression of E-cadherin in MKN45 cells was significantly increased in AEG-1 interference group. These results suggest that digoxin can inhibit the migration and invasion of human gastric cancer MKN45 cells in a dose-dependent manner, which may be associated with digoxin inhibiting the expression of AEG-1 protein. The down-regulation of MMP-9 protein was related to the up-regulation of E-cadherin protein expression.
【作者单位】：漯河医学高等专科学校医学生物工程重点实验室;
【基金】：supported by the Science and Technology Development Project of the Science and Technology Department of Henan Province,China(No.132102310356) the Natural Science Research Project of Luohe Medical College,Henan Province,China(No.2013-S-LMC07)
【分类号】：R735.2

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