CAJ | 학술논문

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，3 d后细胞增殖率较对照组下降57．7％；Wip1基因沉默组TMZ处理5 d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。
목적：연구파향침묵Wip1기인대증민체막서알（ TMZ）억제뇌효질류세포증식작용적영향。방법체외배양인효질류세포주U-87MG，장휴대Wip1기인RNA간우재체적만병독감염U87-MG세포。사용TMZ간예효질류세포，MTT법검측세포적증식，류식세포술Annexin-V （ APC염색）검측세포적조망정황，류식세포술PI염색법검측세포주기。실험수거채용SPSS 16．0연건진행통계학분석。결과 Wip1기인침묵적효질류세포여공재체만병독대조조세포대TMZ적작용대비，3 d후세포증식솔교대조조하강57．7％；Wip1기인침묵조TMZ처리5 d후세포조망솔위15．3％，공재체대조조위5．65％；Wip1기인침묵조세포조망솔명현요고（ P＜0．05）；세포주기결과현시Wip1기인침묵조G2세포위63．0％，이공재체대조조위23．8％，Wip1기인침묵조표현위G2／M기세포명현증다（P＜0．05）。결론파향침묵Wip1기인가이현저증가TMZ대효질류세포증식적억제작용。
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 .