重组人肠三叶因子对结肠癌HT-29细胞移行能力的影响及其机制研究
投稿时间:2014-09-18  修订日期:2014-09-30  点此下载全文
引用本文:李腾,彭曦.重组人肠三叶因子对结肠癌HT-29细胞移行能力的影响及其机制研究[J].医学研究杂志,2015,44(3):37-40
DOI: 10.3969/j.issn.1673-548X.2015.03.011
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作者单位E-mail
李腾 710021 西安医学院护理学院  
彭曦 重庆, 中国人民解放军第三军医大学西南医院烧伤研究所 pxlrmm@163.com 
基金项目:国家自然科学基金面上项目(81372049)
中文摘要:目的 观察重组人肠三叶因子对HT-29细胞移行能力的影响并探讨其作用机制。方法 采用重组表达的人肠三叶因子(rhITF)作为细胞刺激药物, 以传代培养的人结肠癌HT-29细胞株为研究模型。用不同浓度(10、25和50μg/ml) rhITF刺激HT-29细胞, 采用Transwell法观察HT-29细胞移行能力的变化;用50μg/ml rhITF在不同时相点分别刺激HT-29细胞, 分为4、8、12和24h组。通过Western blot法观察黏附蛋白β-catenin、E-cadherin和磷酸化β-catenin的蛋白表达变化。结果 rhITF促细胞移行能力随其浓度的增加而增强, 50μg/ml rhITF组细胞数明显高于阴性对照组、10μg/ml rhITF组和25μg/ml rhITF组, 差异有统计学意义(P<0.01);β-catenin和E-cadherin的蛋白表达均受到rhITF抑制, 与其他组比较, 12h组蛋白表达明显减弱(P<0.05);12h组磷酸化β-catenin 的蛋白表达明显增强(P<0.05)。结论 ITF具有促进HT-29细胞移行的能力, ITF能促使β-catenin磷酸化并抑制β-catenin与E-cadherin的表达。
中文关键词:肠三叶因子  HT-29细胞  细胞移行  黏附分子  β-catenin  E-cadherin
 
Effects of Recombinant Human Intestinal Trefoil Factor on Migration in HT-29 Cells and its Mechanism
Abstract:Objective To observe the effects of recombinant human intestinal trefoil factor(rhITF) on HT-29 cell migration, and explore its possible mechanism. Method Recombinant human intestinal trefoil factor (rhITF) was used as stimulation drugs, and subcultured human colon cancer cell (HT-29) for research model. With different concentrations (10, 25 and 50μg/ml) rhITF stimulate HT-29 cells, the change of cell migration ability was observed by Transwell. At different time points with 50μg/ml rhITF were used to stimulate HT-29 cells, and they were divided into 4h, 8h, 12h and 24h group. The change of β-catenin, E-cadherin and phosphorylated β-catenin were observed by Western blot. Results RhITF could promote HT-29 migration. The ability of cell migration was along with the concentration increase. The number of cell in 50μg/ml rhITF group was more than negative control group and 10μg/ml rhITF group and 25μg/ml rhITF group, and the difference was statistically significant (P<0.01). Protein expression of β-catenin and E-Cadherin were inhibited by rhITF. Compared with normal control group, the protein expression of 12h group decreased obviously (P<0.05); the protein expression of phosphorylation β-catenin in 12h group increased significantly (P<0.05). Conclusion ITF could promote HT-29 cell migration.ITF could promote β-catenin phosphorylation, and inhibit protein expression of β-catenin and E-cadherin.
keywords:ITF  HT-29cells  Cell migration  Adhesion molecule  β-catenin  E-cadherin
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