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(rh ITF)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)rh ITF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 rh ITF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rh ITF促细胞移行能力随其浓度的增加而增强,50μg/ml rh ITF组细胞数明显高于阴性对照组、10μg/ml rh ITF组和25μg/ml rh ITF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rh ITF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。
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 subcul- tured human colon cancer cell （ HT - 29） for research model. With different concentrations （ 10, 25 and 50μg/ml） rhlTF stimulate HT - 29 cells, the change of cell migration ability was observed by Transwell. At different time points with 50μg/ml rhlTF were used to stimu- late 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 rhlTF group was more than negative control group and 10μg/ml rhITF group and 25μg/ml rhlTF 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 13 - catenin phosphorylation, and inhibit protein expression of β- catenin and E - cadherin.
出处
《医学研究杂志》
2015年第3期37-40,共4页
Journal of Medical Research
基金
国家自然科学基金面上项目(81372049)
