论文部分内容阅读
目的观察Survivin基因特异性短发卡RNA(shRNA)对人脑胶质母细胞瘤U251细胞体外生长和细胞凋亡的影响。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,分别采用细胞计数法绘制生长曲线和平板克隆形成实验观察各组细胞的体外生长情况;采用流式细胞术(FCM)定量测定各组细胞的细胞周期和凋亡细胞数量;采用HE染色、Hoechst染色和原位末端标记(TUNEL)方法观察各组细胞形态学特征。结果与U251和U251-P细胞相比,U251-SR细胞生长明显减缓(P<0.01),细胞克隆形成明显减少(P<0.01);U251-SR细胞G1期细胞增多,S期细胞减少,凋亡细胞增加至20.9%,并呈现典型的细胞凋亡形态学改变。结论靶向Survivin基因的特异性shRNA能够在体外明显抑制U251细胞的生长并诱导其发生大量凋亡。
Objective To investigate the effects of Survivin gene-specific short hairpin RNA (shRNA) on the growth and apoptosis of human glioblastoma U251 cells in vitro. Methods U251 cells, U251-SR cells stably transfected with Survivin gene shRNA eukaryotic expression vector pWH1-SR and U251-P cells stably transfected with empty vector pWH1 were used to calculate the cell growth curve and plate clone formation assay The growth of each group of cells was observed in vitro. The cell cycle and the number of apoptotic cells in each group were quantitatively determined by flow cytometry (FCM). The cells in each group were observed by HE staining, Hoechst staining and TUNEL method Morphological characteristics. Results Compared with U251 and U251-P cells, the growth of U251-SR cells was significantly slowed down (P <0.01), and the cell clone formation was significantly reduced (P <0.01). U251-SR cells increased in G1 phase and decreased in S phase The number of dead cells increased to 20.9%, and showed a typical apoptotic morphological changes. Conclusion Specific shRNA targeting Survivin gene can significantly inhibit the growth of U251 cells in vitro and induce a large number of apoptosis.