摘要
背景:肾母细胞瘤过度表达基因(NOV)编码的NOV蛋白是一种胰岛素样生长因子(insulin-likegrowthfactor,IGF)结合蛋白,它对人神经干细胞的增殖和分化有什么影响作用呢?目的:探讨NOV蛋白对人神经干细胞(HNSCs)增殖和分化的影响。设计:以细胞为研究对象,单因素方差分析实验研究。单位:一所军医大学组织学与胚胎学教研室和神经生物学教研室。材料:实验在解放军第三军医大学组织学与胚胎学教研室完成。对象为10～14周人胚大脑皮质培养的HNSCs。干预:NOV基因重组质粒转染COS-7细胞,收集COS-7细胞和NOV基因修饰COS-7细胞的条件培养液(COS-CM和NOV-CM),作用于培养的HNSCs。主要观察指标:3H-TdR掺入液闪检测HNSCs的增殖,免疫细胞化学和流式细胞仪检测HNSCs的分化。结果:COS-CM和NOV-CM均能明显促进HNSCs对3H-TdR的摄入,其中NOV-CM组的1/min比COS-CM组要高(P<0.05),说明NOV-CM中含有促进HNSCs增殖的成分,另外NOV-CM的促细胞增殖作用具有一定的量效关系;免疫细胞化学和流式细胞仪结果显示,在NOV-CM组内有较多的NF-200阳性细胞,在对照组和COS-CM组可见到大量GFAP阳性细胞。结论:NOV蛋白可能具有促进HNSCs增殖和向神经元分化的作用。
BACKGROUND:NOV protein encoded by nephroblastoma overexpression gene(NOV) is IGF(insulin like growth factor) binding protein.What is its impact on human neural stem cell(hNSC) proliferation and differentiation? OBJECTIVE:To investigate the impacts of NOV protein on hNSCs proliferation and differentiation. DESIGN:A single factor analysis of variance experimental study using cells as subjects SETTING:Department of histology and embryology,and department of neurobiology in a military medical university. MATERIALS:Study was conducted in the Department of Histology and Embryology of the Third Military Medical University of Chinese PLA.Subjects were hNSCs cultured from 10 to 14 weeks human embryo cerebral cortex. INTERVENTIONS:COS 7 cells were transfected by NOV gene recombined plasmid.COS 7 cell and COS 7 cell modified by NOV gene conditioned culture media(COS CM and NOV CM) were collected and reacted with the cultured HNSCs. MAIN OUTCOME MEASURES:hNSCs proliferation was detected by 3H TdR scintillation analysis,and hNSCs differentiation was detected by immunocytochemistry and flow cytometer(FCM). RESULTS:Both COS CM and NOV CM could significant promote the intake of 3H TdR by HNSCs,of which the 1/minute of NOV CM group was significantly higher than that of COS CM group(P< 0.05),which indicated that NOV CM contained component that could facilitate hNSCs proliferation, and moreover,there was certain dose effect relationship in NOV CM's facilitation of cellular proliferation.The results of immunocytochemistry and FCM revealed that there were more NF 200 positive cells in NOV CM group,while many glial fibrillary acidic protein positive cells could be seen in COS CM group. CONCLUSION:NOV protein might have facilitative effects on hNSCs proliferation and differentiation into neurons.
出处
《中国临床康复》
CSCD
北大核心
2005年第5期202-204,i004,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30170487
30100173)
全军医学科研"十五"计划青年基金资助项目(01Q097)
重庆市科委应用基础研究项目(2001)~~
