摘要
目的探讨PeroxiredoxinsⅢ(PRDXⅢ)基因对人胶质瘤细胞株U251体外增殖和凋亡的影响。方法将U251细胞分为空白对照组、空载体转染组、无关序列转染组和干扰组。利用脂质体将PRDXⅢ干扰片段转染胶质瘤细胞株U251,RT-PCR及Western blot检测转染后PRDXⅢmRNA及蛋白的表达水平,流式细胞仪(FCM)、DNA ladder、PI染色检测转染后细胞的凋亡状况,四甲基偶氮唑盐(MTT)检测转染后U251细胞的生长抑制率。结果转染后干扰组PRDXⅢmRNA及蛋白的表达水平明显降低,抑制率分别为76.0%和63.9%。DNA ladder显示干扰组出现明显梯形电泳条带。PI染色结果显示干扰组细胞核的染色质高度浓染色,部分细胞核裂解为碎块。空白对照组、空载体转染组、无关序列转染组和干扰组细胞凋亡率分别为(2.26±0.64)%、(2.68±0.74)%、(3.54±0.62)%和(35.90±4.85)%,干扰组细胞凋亡率明显高于其他各组(P=0.00)。干扰组U251细胞生长抑制率为44.40%。结论 PRDXⅢ靶向干扰能够显著抑制U251细胞株增殖,增加其凋亡。
Objective To investigate the effects of Peroxiredoxins Ⅲ（PRDX Ⅲ） gene on proliferation and apoptosis of U251 human glioma cells in vitro.Methods U251 cells were divided into blank control group,empty vector transfection group,unrelated sequence transfection group and interference group.PRDX Ⅲ interference fragment was transfected into U251 glioma cells via liposome.RT-PCR and Western blot were employed to detect the mRNA and protein expression of PRDX Ⅲ in transfected cells,respectively.Flow cytometry（FCM）,DNA ladder and propidium iodide（PI） staining were used to determine the apoptosis of transfected U251 cells.MTT was adopted to measure the rate of growth inhibition of transfected U251 cells.Results In interference group,the mRNA and protein expression of PRDX Ⅲ of cells were reduced significantly after transfection,with inhibitory rates of 76.0% and 63.9%,respectively,DNA ladder showed obvious ladder-shaped electrophoretic bands,and PI staining revealed nuclear chromatin with high staining and some nuclear fragments.Apoptotic rates of cells in blank control group,empty vector transfection group,unrelated sequence transfection group and interference group were（2.26±0.64）%,（2.68±0.74）%,（3.54±0.62）% and（35.90±4.85）%,respectively.Apoptotic rate of cells in interference group was markedly higher than that in other groups（P=0.00）.Growth inhibitory rate of U251 cells in interference group was 44.40%.Conclusion PRDX Ⅲ targeted interference can significantly inhibits proliferation and increases apoptosis of U251 cells.
出处
《重庆医学》
CAS
CSCD
北大核心
2011年第35期3594-3596,F0003,共4页
Chongqing medicine
