RNAi沉默PTTG基因对恶性胶质瘤U251细胞的影响

The effect of PTTG gene silenced by RNAi on inhibition of human glioblastoma U251 cells

目的探讨垂体瘤转化基因(PTTG)siRNA干扰质粒对人脑胶质瘤细胞株U251体外增殖、周期和凋亡的影响。方法利用脂质体将pGenesil-2 PTTG siRNA干扰质粒转染U251细胞,通过MTT法观察转染后24h、48h和72h细胞增殖的变化、流式细胞术PI单染观察转染后48h细胞凋亡和细胞周期的变化。结果pGenesil-2PTTG siRNA干扰质粒转染U251细胞48h后,PTTG mRNA和蛋白的抑制率分别约为69%和71%;阻滞U251细胞由G1期进入S期,抑制细胞的生长,增加了细胞的凋亡。结论PTTG siRNA表达载体可以特异高效地抑制胶质瘤U251细胞PTTG基因的表达,从而调控肿瘤细胞周期,抑制肿瘤细胞增殖。

Objective To observe the effects of pituitary tumor transforming gene （PTrG） siRNA specific expression vector on U251 glioma cells proliferation, cycle and apoptosis. Methods The PTrG siRNA expression vector and empty expression vector were transfected into U251 cells by liposome 2000 to observe cell proliferation 24, 48 and 72 h later by MTT method and apoptosis 48 h after transfection and cell cycle by flow cytometry PI simple stain. Results The PTTG siRNA expression vector significantly decreased the expression levels of PTTG mRNA and protein, and the inhibition rates were 69% and 71% respectively. It also inhibitod the transition from G1 phase to S phase and the growth of U251 cells and promoted apoptosis. Conclusions PTTG specific siRNA expression vector could specifically and efficiently inhibit the expression of PTTG gene to regulate cell cycle and inhibit cell proliferation in U251 glioma cells.