Notehl对人脐带问充质干细胞增殖、黏附及迁移影响的体外研究

目的探讨Notchl基因的重组腺病毒转染对人脐带间充质干细胞（humanumbilicalcordmesenchymalstemcells，hUCMSCs）增殖、黏附及迁移能力的影响。方法按照不同腺病毒感染hUCMSCs分为实验组（Ad-Notchl）、阴性对照组（Ad—GFP）与空白对照组。以最佳MOI转染hUCMSCs，观察细胞形态变化以及GFP蛋白的表达。比较3组细胞增殖能力、细胞周期、黏附能力及迁移能力。WesternBolt检测3组细胞Notchl蛋白的表达水平。结果荧光下观察腺病毒成功感染细胞，阴性对照组和实验组均出现了GFP的表达，且随培养时间的延长表达量增高，可见光下观察实验组细胞增殖速度较快。实验组细胞的增殖能力（48、72、96h）、黏附能力（12、16、20h）及迁移能力均较阴性对照组及空白对照组增强（P〈0．05）。WesternBolt检测实验组细胞Notchl蛋白的表达水平显著高于阴性对照组和空白对照组（P〈0．05）。结论Notchl能明显促进hUCMSCs的增殖、黏附和迁移。

Effect of Notchl on Proliferation, Adhesion and Migration of Human Umbilical Cord Mesenchymal Stem Cells in Vitro

Objective To discuss the effect of transfection of adenovirus recombinant in Notchl gene on the pro- liferation, adhesion and migration of human umbilical cord mesenchymal stem cells （hUCMSCs）. Methods The hUC- MSCs were divided into experimental group （Ad-Notehl group） , negative control group （Ad-GFP group） and control group according to the different adenovirus infections. HUCMSCs were transfected by the optimal multiplicity of infection （MOI） , and changes in cell morphology and the expression of green fluorescent protein （GFP） were observed. The cell proliferation ability, cell cycle, adhesion ability and migration ability in the three groups were compared. The expression level of Notchl protein in each group was detected by Westeru Bolt method. Results Adenovirus successfully infected cells under fluorescence, and the GFP expression was observed in control and Ad-Notchl groups, and the expression level was higher with prolate cultivation time; so the speed of cell multiplication was quick under visible light in experimental group. Abilities of proliferation （48 h, 72 h and 96 h）, adhesion （12 h, 16 h and 20 h） and migration of ceils in experi- mental group were significantly enhanced, compared with those in negative control group and control group （ P 〈 0.05）. The expression level of Notchl protein in experimental group was significantly higher than those in negative control group and control group by Western Bolt method （P 〈 0.05）. Conclusion Notchl can remarkably promote the ability of pro- liferation, adhesion and migration in hUCMSCs.