siRNA抑制hTERT对脑胶质瘤T98G细胞增殖及周期的影响  

Effect of siRNA's inhibition of hTERT on T98G glioma cell proliferation and cycle

作　　者：王拓[1] 王举波[1] 李超[2] 董丹凤[3] 王茂德[1] 

机构地区：[1]西安交通大学医学院第一附属医院神经外科,陕西西安710061 [2]西安交通大学医学院公共卫生学系,陕西西安710061 [3]西安交通大学医学院第一附属医院肿瘤内科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2012年第5期555-559,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的研究人端粒酶逆转录酶被抑制后胶质瘤细胞系体外增殖及周期的变化,以期寻找控制胶质瘤细胞恶性行为的有效途径。方法人工合成小干扰RNA(siRNA)序列,并将其通过lip2000转入体外培养的T98G胶质瘤细胞系中,荧光显微镜观察转染效率,免疫印记检测蛋白表达情况,MTT法检测细胞体外增殖表现,绘制生长曲线图,流式细胞仪检测细胞周期的变化。结果成功转染siRNA,并有效抑制人端粒酶逆转录酶的活性,免疫印记验证干扰效果可靠,体外可抑制胶质瘤细胞的增殖,与对照组比较其抑制效率的差异有统计学意义(P<0.05)。结论体外条件下,可以通过siRNA有效抑制人端粒酶逆转录酶,从而有效抑制胶质瘤细胞的体外增殖,使胶质瘤细胞周期发生变化。这可能为胶质瘤的治疗带来新的启示。Objective To study the proliferation and cycle change of glioma cell lines in vitro after human telomerase reverse transcriptase(hTERT) is effectively inhibited so as to seek an effective way to control malignant behavior of glioma cells.Methods Effective small interfering RNA sequence was made,and then transferred into cultured T98G glioma cell lines through the lip2000.The transfection efficiency was observed by fluorescence microscopy.Western blot was used to verify the protein expression;MTT assay was used to observe cell lines proliferation;flow cytometry was used to detect the cell cycle changes.Results Small interference RNA was transfected successfully.Transfected siRNA could inhibit hTERT effectively,and then inhibit the proliferation of cell lines and change the cell cycle in vitro.The inhibition rate differed significantly from that of the control group(P<0.05).Conclusion Small RNA is an effective way for gene interference.It can silence hTERT,thus inhibiting the proliferation of T98G cell lines and changing the cell cycle.Therefore,it may be a hopeful way of curing glioma.

关 键 词：胶质瘤 小干扰RNA 转染 人端粒酶逆转录酶 细胞增殖 细胞周期 

分 类 号：R739.4[医药卫生—肿瘤]