摘要
目的研究血管细胞粘附分子-1(VCAM-1)在骨髓间充质干细胞(BMSCs)中的表达及胶质瘤细胞对其表达的影响,探讨VCAM-1在BMSCs向胶质瘤趋化迁移中的作用。方法骨髓间充质干细胞分离自4-6周龄SD大鼠,采用全骨髓贴壁法原代培养。利用大鼠C6胶质瘤细胞条件培养基孵育BMSCs24h,利用免疫荧光及RT-PCR检测BMSCs的VCAM-1表达变化情况。利用Transwell构建BMSCs体外迁移模型,观察BMSCs向大鼠C6胶质瘤细胞的趋化迁移情况。然后在Transwell上室加入VCAM-1特异性阻断单克隆抗体,观察阻断VCAM-1前后BMSCs向胶质瘤细胞迁移的变化。结果 Transwell体外迁移结果显示BMSCs在体外具有向胶质瘤细胞条件培养上清迁移的能力。免疫荧光及RT-PCR结果显示,BMSCs较弱表达VCAM-1;加入C6胶质瘤细胞条件培养基后,VCAM-1的蛋白及mRNA表达增强;加入特异性抗体阻断VCAM-1后,BMSCs向胶质瘤细胞的迁移受到抑制。结论胶质瘤细胞可在体外通过上调BMSCs表达VCAM-1而促进其向胶质瘤趋化迁移。
Objective The aims of this research were to observe the expression of vascular cell adhesion molecule-1（VCAM-1） in bone marrow-derived mesenchymal stem cells（BMSCs） as well as the change of VCAM-1 expression by glioma cells and to investigate the role of VCAM-1 in the tropism of BMSCs toward glioma. Methods BMSCs were isolated from 4-6 weeks old female Sprague-Dawley rats and cultured by means of cell attachment to culture plate. BMSCs were treated with the conditioned culture of C6 glioma cells and the expression pattern was checked by immunofluorescence and RT-PCR. The migration of BMSCs toward glioma cells was investigated with Transwell migration assay. The blocking monoclonal antibody was added into the upper chamber to neutralize the function of VCAM-1 and the influence to migration was investigated. Results BMSCs displayed the capacity of migrating toward glioma in vitro. The results of immunofluorescence and RT-PCR showed that VCAM-1 was expressed in BMSCs and upregulated by glioma ceils. The migration of BMSCs toward glioma was inhibited significantly with the blocking of VCAM-1. Conclusion Glioma cells promote the migration through up-regulating the expression of VCAM-1 in BMSCs.
出处
《解剖科学进展》
CAS
2013年第3期201-205,共5页
Progress of Anatomical Sciences
基金
国家自然科学基金(No.30901781
81172197
81072056)
辽宁省博士启动基金(No.20091107)资助项目
