载体介导的SIRNA抑制B7-H1在星形胶质细胞上的表达  

siRNA vector silencing B7-H1 expression in astrocyte

作　　者：熊加祥[1] 许雪青[1] 白云[1] 陈雪丹[1] 王璞[1] 宋敏[1] 

机构地区：[1]第三军医大学基础医学部分子遗传学教研室

出　　处：《第三军医大学学报》2006年第4期298-300,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金海外青年学者合作研究基金资助项目(30228018);第三军医大学校管课题(200503)~~

摘　　要：目的构建SIRNA稳定表达载体,抑制B7-H1分子在星形胶质细胞上的表达,为研究大脑内B7-H1分子的功能提供有效实验工具。方法设计合成带有双向启动子具有转录功能性SIRNA载体,转入小鼠星形胶质细胞株CRL-2541内,干扰免疫分子B7-H1的表达,利用荧光载体N1平行观察转染效果,RT-PCR方法检测B7-H1的MRNA表达,免疫组化检测蛋白表达水平,以观察干扰效率。结果星形胶质细胞株CRL-2541经RNAI沉默后B7-H1MRNA表达下调,免疫组化发现其蛋白水平下降。结论成功建立具有双向启动子的SIRNA载体,并在星形胶质细胞中抑制B7-H1的表达。Objective To silence the expression of B7-H1 in astrocyte by stable expression vector containing siRNA, providing experiment instrument for researching function of B7-H1 in brain. Methods The siRNA vector possessing bidirectional promoter and transcription function was designed and synthesized, then transfected into mouse astrocyte CRL-2541 to silence the expression of B7-H1. The effect of transfection was observed with fluorescence N1 vector. The expression of BT-H1 mRNA was detected by RT-PCR and BT-H1 protein by immunohistochemistry to study efficiency of interference. Results The expression of B7-H1 mRNA and protein in astrocyte line CRL-2541 was decreased after RNAi. Conclusion SiRNA vector with bidirectional promoter was constructed successfully and can silence the expression of B7-H1 in astrocyte.

关 键 词：RNAI B7-H1 星形胶质细胞 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学] R394.33[医药卫生—基础医学]