摘要
目的:观察针对Survivin基因的短发卡RNA(shRNA)对人脑胶质母细胞瘤U251细胞在体外凋亡的影响。方法:对U251细胞,稳定转染Survivin基因shRNA真核表达载体pWH1-SR的U251-SR细胞,以及稳定转染空载体pWH1的U251-P细胞,分别采用相差显微镜、HE染色、Hoechst染色、TUNEL染色以及透射电镜观察各组细胞的形态学特征;采用流式细胞术(FCM)定量测定各组细胞的凋亡细胞数量。结果:相差显微镜、HE染色、Hoechst染色、TUNEL染色以及透射电镜观察显示,U251-SR凋亡细胞明显增多,并呈现典型的细胞凋亡形态学改变;FCM定量分析凋亡细胞数量显示,与U251(2.1%)和U251-P(2.7%)相比,U251-SR凋亡细胞增加约6倍,达14.4%。结论:针对Survivin基因的shRNA能够在体外诱导U251细胞发生大量凋亡。
Objective： To observe the effect of short hairpin RNA （shRNA） targeting the survivin gene on apoptosis of human brain glioblastoma U251 cells in vitro. Methods： For the U251 cells, U251-SR cells stably transfected with the shRNA eukaryotic expression vector pWH1-SR targeting the survivin gene, and U251-P cells stably transfected with the empty pWHI vector, phase contrast mi- croscopy, H＆E staining, Hoechst staining, TUNEL staining and transmission electron microscopy were used to observe the morphologic characteristics of the cells in each group. Flow cytometry （FCM） was used to determine the number of apoptotic cells in each group. Results： The results of phase contrast microscopy, H＆E stairiing, Hoechst staining, TUNEL staining and transmission electron microscopy showed that the percentage of apoptotic cells among the U251-SR ceils significantly increased, pro- ducing morphologic changes typical of apoptosis. The FCM quantitative analysis showed that the U251- SR apoptotic cells amounted to 14.4% of the total with the U251 （2.1%） and U251-P （2.7%） cells. induce apoptosis of U251 cells in vitro. population, an increase of about 6-fold, compared Conclusion： shRNA targeting the survivin gene can
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2007年第4期186-189,共4页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金资助(编号:39970752)
