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| Snail基因修饰对骨髓基质干细胞骨架结构稳定作用及促迁移和对无血清培养诱导细胞凋亡的保护作用研究 | |
| 引用本文： | 查运红,梅元武,毛玲,贺杰峰,殷涛,张立,王全胜. Snail基因修饰对骨髓基质干细胞骨架结构稳定作用及促迁移和对无血清培养诱导细胞凋亡的保护作用研究[J]. 生物工程学报, 2007, 23(4): 645-651 |
| 作者姓名： | 查运红 梅元武 毛玲 贺杰峰 殷涛 张立 王全胜 |
| 作者单位： | 1. 华中科技大学同济医学院附属协和医院神经内科,武汉,430022 2. 华中科技大学同济医学院附属同济医院普外科,武汉,430030 3. 华中科技大学同济医学院基础医学院病理生理学系,武汉,430030 4. 华中科技大学同济医学院附属协和医院肾内科,武汉,430022 |
| 摘 要： | 转录因子Snail是调控肿瘤细胞迁徙转移的重要调控分子,基于干细胞与肿瘤细胞分子机制的重叠性,提出通过借鉴肿瘤细胞迁移的相关机制以用于提高骨髓基质干细胞向缺氧受损组织迁移能力的假设和研究思路,探讨Snail基因在人骨髓基质干细胞(MSCs)中的转染和表达情况,及转染后对基质干细胞促迁移作用、骨架结构的稳定作用及对无血清培养诱导细胞凋亡的保护作用。密度梯度离心法及细胞体外培养分离纯化人骨髓MSCs,脂质体法将重组表达载体pCAGGSneo-Snail-HA转染MSCs,G418筛选稳定表达,流式细胞仪检测MSCs表面抗原,采用免疫荧光染色技术检测转染后MSCs报告基因HA及目的基因Snail表达,Transwell细胞迁移实验和Western-blot评估细胞迁移能力和检测有关细胞信号转导通路分子水平变化,荧光染色分析细胞骨架,Sub-G1凋亡峰流式细胞仪检测细胞凋亡率并评估细胞抗凋亡能力。经流式细胞仪选择检测分离纯化扩增MSCs表面分子特点为CD34(-)/CD29( ),Snail及报告基因在转染后MSCs呈阳性表达,Snail质粒转染MSCs(MSCs-Sna)较对照空质粒转染MSCs(MSCs-neo)细胞迁移率增加(P<0.05),PI-3K信号通路特异性抑制剂Wortmannin能显著抑制此迁移率的增加,无血清培养72h后,MSCs-Sna凋亡率较MSCs-neo低(P<0.05)。经Snail基因转染,MSCs迁移能力、骨架结构的稳定性及在无血清培养环境中抗凋亡能力增加。 |
| 关 键 词： | 骨髓基质干细胞 Snail转录因子 肿瘤细胞 迁移 细胞骨架 凋亡 |
| 文章编号： | 1000-3061（2007）04-0645-07 |
| 修稿时间： | 2006-08-22 |
The Advantages for Snail Expression to Promote Cell Migration and Induce Actin Reorganization and to Protect Against the Serum-deprivation-triggered Apoptosis of Bone Marrow Stem Cells |
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| ZHA Yun-Hong,MEI Yuan-Wu,MAO Ling,HE Jie-Feng,YIN Tao,ZHANG Li and WANG Quan-Sheng. The Advantages for Snail Expression to Promote Cell Migration and Induce Actin Reorganization and to Protect Against the Serum-deprivation-triggered Apoptosis of Bone Marrow Stem Cells[J]. Chinese journal of biotechnology, 2007, 23(4): 645-651 | |
| Authors: | ZHA Yun-Hong MEI Yuan-Wu MAO Ling HE Jie-Feng YIN Tao ZHANG Li WANG Quan-Sheng |
| Affiliation: | Department of Neurology, Union Hospital , Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China Department of Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China Department of Pathophysiology , Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China Department of Nephrology , Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China |
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