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地高辛通過下調(diào)AEG-1表達(dá)抑制人胃癌MKN45細(xì)胞遷移和侵襲

發(fā)布時(shí)間:2018-02-28 16:18

  本文關(guān)鍵詞: 地高辛 星形細(xì)胞上調(diào)基因- 遷移 侵襲 胃癌 MKN細(xì)胞 出處:《生理學(xué)報(bào)》2017年02期  論文類型:期刊論文


【摘要】:本研究旨在觀察地高辛對(duì)人胃癌MKN45細(xì)胞遷移和侵襲能力的影響,并探討其分子機(jī)制。取人胃癌MKN45細(xì)胞作為研究對(duì)象,采用Transwell法檢測(cè)細(xì)胞遷移和侵襲能力,通過脂質(zhì)體轉(zhuǎn)染法將星形細(xì)胞上調(diào)基因-1(astrocyte elevated gene-1,AEG-1)sh RNA干擾質(zhì)粒轉(zhuǎn)染MKN45細(xì)胞以構(gòu)建低表達(dá)AEG-1的細(xì)胞株,利用Western blot法檢測(cè)基質(zhì)金屬蛋白酶-9(matrix metalloproteinase-9,MMP-9)、E-鈣粘蛋白(E-cadherin)和AEG-1等蛋白的表達(dá)變化。結(jié)果顯示,地高辛可使MKN45細(xì)胞遷移率和侵襲率均顯著下降(P0.05),下調(diào)MMP-9和AEG-1蛋白表達(dá)水平(P0.05)以及上調(diào)E-cadherin蛋白表達(dá)水平(P0.05),上述作用均具有劑量依賴性。經(jīng)sh RNA干擾AEG-1基因表達(dá)后,MKN45細(xì)胞中AEG-1蛋白表達(dá)水平顯著下降(P0.05);同時(shí)AEG-1干擾組MKN45細(xì)胞中E-cadherin蛋白表達(dá)水平顯著升高(P0.05),細(xì)胞遷移率、侵襲率和MMP-9蛋白表達(dá)水平均顯著下降(P0.05)。上述結(jié)果提示,地高辛可在體外劑量依賴性地抑制人胃癌MKN45細(xì)胞遷移和侵襲,這可能與地高辛抑制AEG-1蛋白表達(dá),繼而下調(diào)MMP-9蛋白表達(dá)和上調(diào)E-cadherin蛋白表達(dá)有關(guān)。
[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.
【作者單位】: 漯河醫(yī)學(xué)高等?茖W(xué)校醫(yī)學(xué)生物工程重點(diǎn)實(shí)驗(yàn)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)
【分類號(hào)】:R735.2

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