地高辛通过下调AEG-1表达抑制人胃癌MKN45细胞迁移和侵袭  

Digoxin inhibits migration and invasion of human gastric carcinoma MKN45 cells through down-regulation of AEG-1

作　　者：崔明辰[1] 王兴红[1] 袁磊[1] 熊均平[1] CUI Ming-Chen WANG Xing-Hong YUAN Lei XIONG Jun-Ping(Key Laboratory of Medical Bioengineering, Luohe Medical College, Luohe 462002, Chin)

机构地区：[1]漯河医学高等专科学校医学生物工程重点实验室,漯河462002

出　　处：《生理学报》2017年第2期189-195,共7页Acta Physiologica Sinica

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)

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(P<0.05),下调MMP-9和AEG-1蛋白表达水平(P<0.05)以及上调E-cadherin蛋白表达水平(P<0.05),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(P<0.05);同时AEG-1干扰组MKN45细胞中E-cadherin蛋白表达水平显著升高(P<0.05),细胞迁移率、侵袭率和MMP-9蛋白表达水平均显著下降(P<0.05)。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。This study was designed to investigate the effect of digoxin on migration and invasion of human gastric carcinoma MKN45 cells and its possible mechanism. MKN45 cells were treated with different concentrations of digoxin for 24 h. The shRNA-AEG-1 plasmid was transfected into MKN45 cells via lipofectamine to block the expression of astrocyte elevated gene-1 （AEG-1）. Western blot was used to analyze the protein levels of matrix metalloproteinase-9 （MMP-9）, E-cadherin and AEG-1. The result showed that digoxin reduced the abilities of migration and invasion （P 〈 0.05）, up-regulated the protein level of E-cadherin （P 〈 0.05）, and down-regulated the protein levels of MMP-9 and AEG-1 （P 〈 0.05） in MKN45 cells in a dose-dependent manner. Compared with shControl group, shAGE-1 group showed inhibited cellular migration and invasion, higher expression level of E-cadherin, and lower expression levels of MMP-9 and AEG-1. These results suggest that digoxin suppresses the migration and invasion of human gastric carcinoma MKN45 cells in a dose-dependent manner through inhibiting the expression ofAEG-1, and then resulting in the up-regula- tion of the protein expression of E-cadherin and the down-regulation of the protein expression of MMP-9.

关 键 词：地高辛 星形细胞上调基因-1 迁移 侵袭 胃癌 MKN45细胞 

分 类 号：R735.2[医药卫生—肿瘤]