摘要
目的观察短发卡RNA(short hairpin RNA,shRNA)沉默Survivin基因对人脑胶质瘤细胞U87增殖、凋亡等生物学特性的影响。方法通过脂质体LipofectamineTM 2000对胶质瘤细胞株U87转染Survivin shRNA干扰质粒(Suvrivin shRNA-i),以及对照无意义质粒(Suvrivin shRNA-nonsense),应用RT-PCR、Western blot检测转染前后Survivin mRNA和蛋白表达,流式细胞术测定各组细胞的细胞周期及TUNEL测定细胞凋亡形态。结果转染Survivin shRNA-i组的Survivin mRNA及蛋白表达水平显著低于对照组(P<0.05)。与对照组相比较,转染Survivin shRNA-i后细胞周期G_2/M期阻滞显著,细胞凋亡率升高(P<0.05)。结论靶向Survivin基因的特异性shRNA能够阻制U87细胞的生长并诱导其凋亡。
Objective To observe the influence of specific short hairpin RNA （shRNA） targeting Survivin gene on proliferation and apoptosis of human brain glioma U87 cell line. Methods Survivin shRNA interfering（ shRNA -i）and nonsense shRNA were transfected respectively into U87 by Lipofectamine 2000. Expression of mRNA and protein of Survivin was detected before and after transfection by RT - PCR and Western blot. Flow cytometry （FCM） was adopted to analyze the cell cycle and quantity of apoptotic cells in each group. TUNEL method was used to observe the morphologic characteristics of the cells in each group. Results The expression of Survivin mRNA and protein level was lower in U87 - shRNA - i group than other ones （ P 〈 0.05 ）. The cell proliferation was slowed down, cell cycle was arrested in G2/M and apoptosis rate was increased. Conclusion The specific shRNA targeting Survivin gene can significantly inhibit the growth and induce the apoptosis of U87 cells in vitro.
出处
《河北医科大学学报》
CAS
2011年第12期1392-1395,F0003,共5页
Journal of Hebei Medical University
