摘要
目的研究靶向沉默Wip1基因对增敏替莫唑胺(TMZ)抑制脑胶质瘤细胞增殖作用的影响。方法体外培养人胶质瘤细胞株U-87MG,将携带Wip1基因RNA干扰载体的慢病毒感染U87-MG细胞。使用TMZ干预胶质瘤细胞,MTT法检测细胞的增殖,流式细胞术Annexin-V(APC染色)检测细胞的凋亡情况,流式细胞术PI染色法检测细胞周期。实验数据采用SPSS 16.0软件进行统计学分析。结果 Wip1基因沉默的胶质瘤细胞与空载体慢病毒对照组细胞对TMZ的作用对比,3d后细胞增殖率较对照组下降57.7%;Wip1基因沉默组TMZ处理5d后细胞凋亡率为15.3%,空载体对照组为5.65%;Wip1基因沉默组细胞凋亡率明显要高(P<0.05);细胞周期结果显示Wip1基因沉默组G2细胞为63.0%,而空载体对照组为23.8%,Wip1基因沉默组表现为G2/M期细胞明显增多(P<0.05)。结论靶向沉默Wip1基因可以显著增加TMZ对胶质瘤细胞增殖的抑制作用。
Objective To study the effects of targeted silencing of Wip 1 gene expression in combination with Temozolomide （ TMZ） on brain glioma cells.Methods Glioma U-87MG cells were cultured and infected with Wip 1 RNAi lentiviral vector .Cells infected with no significant lentiviral vector were used as negative controls .Then the cells were treated with TMZ .The proliferation activities of cells with Wip1 silencing in combination with TMZ were detected by MTT .Cell apoptosis was determined by flow cytometry Annexin V-APC labeling method .Cell cycle distributions were analyzed by flow cytometry PI staining .SPSS 16.0 software was used to analyze the significant difference.Results U-87MG cells with Wip1 silencing treated with TMZ had reduced proliferation ability compared with negative control cells and mock cells .Wip1 silencing in combination with TMZ reduced cell proliferation by 57.7%4 d after the treatments.The apoptotic cells accounted for 15. 3%of all the U-87 MG cells 5 d after the treatments , while accounting for 5 .65% in the control cells.The cells were more apoptotic when Wip 1 silencing in combination with TMZ .Cell cycle distributions analyzed by flow cytometry showed increased G 2/M phase,which accounted for 63.0%cells compared with 23.8%in negative control group（P〈0.05）.Conclusion Targeted silencing of Wip1 gene expression in combination with Temozolomide can prevent proliferation and induce apoptosis of glioma cell .
出处
《临床神经外科杂志》
CAS
2015年第1期1-4,共4页
Journal of Clinical Neurosurgery
基金
国家自然科学基金(81201995)
广东省医学科研基金(A2012009)
