靶向存活素基因短发卡RNA对人脑胶质母细胞瘤U251细胞在裸鼠体内生长的影响  

Inhibition of growth and angiogenesis of U251 cell xenograft in vivo by short hairpin RNA targeting survivin gene

作　　者：甄海宁[1] 章翔[1] 师长宏[2] 杨彤涛[3] 付洛安[1] 章薇[1] 王西玲[1] 高大宽[1] 胡世颉[1] 宋蕾[1] 

机构地区：[1]第四军医大学西京医院全军神经外科研究所,西安710033 [2]基础部实验动物中心 [3]第四军医大学唐都医院全军骨科中心

出　　处：《中华外科杂志》2006年第18期1270-1274,共5页Chinese Journal of Surgery

基　　金：国家自然科学基金(39970752)

摘　　要：目的观察靶向存活素基因的特异性短发卡 RNA(shRNA)对人脑胶质母细胞瘤 U251细胞裸鼠体内肿瘤生长和血管形成的影响。方法将 U251细胞、稳定转染存活素基因 shRNA 真核表达载体 pWH1-SR 的 U251细胞(U251-SR 细胞)、稳定转染空载体 pWH1的 U251细胞(U251-P 细胞),分别接种于15只裸鼠背部皮下,每组5只,观测肿瘤生长情况。采用免疫组化 SABC 方法检测存活素、增殖细胞核抗原(PCNA)以及第八因子相关抗原(FⅧRAg)在各组肿瘤标本中的表达;采用TUNEL 方法检测凋亡细胞,分别计算各组肿瘤标本的增殖指数(PJ)、凋亡指数(AI)以及微血管密度(MVD)。结果与 U251、U251-P 组相比,U251-SR 组裸鼠肿瘤形成时间延迟,肿瘤生长缓慢,肿瘤体积及重量均明显减小(P 均<0.01);肿瘤标本存活素蛋白表达明显下调;PI 和 MVD 明显减少,AI 明显升高(P 均<0.01)。结论靶向存活素基因的 shRNA 能够在裸鼠体内明显抑制 U251细胞的肿瘤生长和血管形成。Objective To observe the impact of specific short hairpin RNA （shRNA） targeting survivin gene on tumorigenesis and angiogenesis of human brain glioblastoma U251 cells in vivo of nude mice. Methods U251 cells, U251-SR cells transfected stably with shRNA eukaryotic expression vector pWH1-SR targeting survivin gene, and U251-P cells transfected stably with blank pWH1 vector, were inoculated respectively into subcutaneous tissue in flank of 15 nude mice（ each group 5 mice） , and the tumor growth status was observed and measured. Protein expressions of survivin, proliferating cell nuclear antigen （PCNA） and factor Ⅷ related antigen （F Ⅷ RAg） were investigated by immunohistochemistry SABC method, apoptotic cells were screened by TUNEL method, furthermore proliferative index （ PI）, apoptotic index （AI） and microvessel density （MVD） were measured respectively in each group of tumor specimens. Results Comparing with those in U251 and U251-P groups, in U251-SR group, the tumorigenesis time delayed, tumor grew slowly, both tumor volume and tumor weight decreased significantly （P 〈 0. 01 for both）; Survivin protein expression was down-regnlated markedly; PI and MVD decreased significantly, whereas AI increased remarkably （ P 〈 0. 01 for all ）. Conclusions The specific shRNA targeting survivin gene can inhibit significantly tumorigenesis and angiogenesis of U251 cells in vivo.

关 键 词：胶质瘤 存活素蛋白 小鼠 RNA干涉 短发卡RNA 

分 类 号：R739.4[医药卫生—肿瘤]