论文部分内容阅读
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 To investigate the expression of vascular cell adhesion molecule-1 (VCAM-1) in bone marrow mesenchymal stem cells (BMSCs) and the effect of VCAM-1 on the expression of VCAM-1 in glioma chemotaxis In the role. Methods Bone marrow mesenchymal stem cells were isolated from 4-6-week-old Sprague-Dawley rats and were primary cultured with whole bone marrow adherent method. BMSCs were incubated with rat C6 glioma conditioned medium for 24 h. The expression of VCAM-1 in BMSCs was detected by immunofluorescence and RT-PCR. Transwell was used to construct the in vitro migration model of BMSCs to observe the chemotactic migration of BMSCs into rat C6 glioma cells. Then, VCAM-1 specific blocking monoclonal antibody was added to the upper chamber of Transwell to observe the change of BMSCs migration to glioma cells before and after blocking VCAM-1. Results Transwell migration in vitro showed that BMSCs had the ability to migrate to conditioned medium of glioma cells in vitro. The results of immunofluorescence and RT-PCR showed that VCAM-1 was weakly expressed in BMSCs, and the expression of VCAM-1 protein and mRNA was increased after adding C6 glioma conditioned medium. After the specific antibody was blocked by VCAM-1, BMSCs Migration to glioma cells is inhibited. Conclusion Glioma cells can promote the chemotactic migration of glioma cells by up-regulating the expression of VCAM-1 in BMSCs.