摘要
目的 检测肾母细胞瘤过度表达基因(nephroblastomaoverexpressedgene ,nov)所编码产物(NOV蛋白)转染嗅鞘细胞(olfactoryensheathingcells ,OECs)后在OECs中的表达以及活性。方法 本研究用nov转染原代培养大鼠OECs ,用G418筛选,经过Westernblot检测c myc NOV的表达以及通过背根神经节(dorsalrootganglion ,DRG)与NOV转染后OECs联合培养来对表达的nov活性进行鉴定。结果 ①Western blot结果显示,转染了NOVcDNA的OECs总蛋白c Myc抗体检测可见约2 8×10 3 条带,即c myc NOV融合蛋白。未转染质粒的OECs总蛋白做对照,结果为阴性,无条带出现。②与nov修饰的OECs联合培养DRG有许多DRG细胞向外迁移,细胞折光性强,突起较长而纤细,并形成复杂的网络。未修饰的OECs组只有少量的细胞迁移和突起生长,细胞迁移和突起生长都很局限。空白对照组DRG仅有细胞迁移。结论 nov转染OECs后,并能促进与OECs联合培养的DRG细胞的迁移生长,表明产生了具有促神经细胞生长活性的蛋白。
s: Objective To study the effect of NOV in the condition medium of nov genetically modified olfactory ensheathing cells (OECs) on dorsal root ganglion (DRG). Methods Chicken E8 DRG were co-cultured with OECs isolated from rat P7 olfactory bulbs. After transfected with nov gene, modified OECs were screened by G418, and c-myc-NOV was confirmed by Western blot and the effect of NOV was tested by coculture with DRG. Results (1) Western blot showed a clear band at 28×10 3, which was c-myc-NOV, while there was no band in the negative control. (2) The DRG co-cultured with nov-modified OECs extended many long neurites that formed a complex network, and just a few cell migrations and neurite outgrowth could be seen when co-cultured with normal OECs. In the blank group, those phenomena could be seen only by chance. Conclusion nov has been transfected into OECs successfully and NOV can promote neurite outgrowth of DRG cells, suggesting that an active NOV is expressed.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2005年第7期596-599,共4页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 30 170 4 87
30 370 4 6 9)
全军医学科研"十五"计划面上项目 ( 0 1MA16 8)
第三军医大学科研基金资助 ( 2 0 0 3)~~
关键词
肾母细胞瘤过度表达基因
嗅鞘细胞
细胞培养
背根神经节
nephroblastoma overexpressed gene
olfactory ensheathing cell
cell culture
dorsal root ganglion
