RNA干扰沉默CXCR4基因对Jurkat细胞周期和凋亡的影响  

Effects of CXCR4 Silence Induced by RNA Interference on Cell Cycle Distribution and Apoptosis of Jurkat Cells

作　　者：王艳[1] 刘晓日[2] 谭艳芳[1] 殷小成[1] 

机构地区：[1]南华大学附属第一医院儿科,湖南衡阳421001 [2]南华大学附属南华医院儿科,湖南衡阳421001

出　　处：《中国实验血液学杂志》2010年第3期625-628,共4页Journal of Experimental Hematology

基　　金：衡阳市2007年科学技术发展计划项目(编号2007KS35)

摘　　要：本文研究探讨下调CXCR4的表达对人T-ALL Jurkat细胞周期和凋亡的影响。设计合成CXCR4的特异性siRNA,采用脂质体转染Jurkat细胞株。用RT-PCR检测siRNA对细胞内CXCR4基因表达的影响,用流式细胞术检测细胞周期分布和细胞的凋亡情况。结果表明:成功构建了CXCR4的特异性siRNA;与空白对照组相比,转染CXCR4 siRNA的Jurkat细胞的CXCR4 mRNA表达水平明显下降(56.9%±1.4% vs 68.3%±2.4%),G0/G1期细胞比例增加(35.8%±1.9% vs 18.1%±1.2%),G2/M期和S期细胞比例相应下降(19.8%±1.7% vs 27.2%±1.5%;44.4%±2.1% vs 54.7%±2.8%),凋亡细胞率明显增高(20.9%±2.0% vs 3.13%±0.9%)。结论:CXCR4的特异性siRNA能够有效下调CXCR4基因表达,诱导Jurkat细胞凋亡和细胞周期阻滞,从而抑制细胞的增殖。This study was aimed to investigate the effect of down-regulating the CXCR4 expression on cell cycle and cell apoptosis of human T-ALL Jurkat cells. The CXCR4 specific siRNA plasmid vector was constructed and then transfected into the cultrued Jurkat cell line by DMRIE-C. The expression of CXCR4 mRNA was detected by RT-PCR, the cell distribution in cell cycle and cell apoptosis were determined by flow cytometry. The experiments were divided into 3 groups： group A （blank control）, group B （ non-silencing dsRNA as negative control ） and group C （ CXCR4 siRNA）. The results showed that the expression level of CXCR4 mRNA in Jurkat cells transfected with CXCR4 siRNA （ group C） decreased and cell proportion in G0/G1 phase increased as compared with group A （ 56.9% ± 1.4% vs 68. 3 % ± 2.4% and 35.8% ± 1.9% vs 18.1% ± 1.2% respectively） （p 〈 0.01 ）, cell proportion in G2/M and S phase decreased as compared with group A （19. 8% ± 1.7%, 44.4% ±2. 1% vs 27. 2% ± 1.5%, 54. 7% ±2. 8% respectively） （p 〈0.01 ）. The apoptosis rate of Jurkat cells in group C increased as compared with group A （20.9% ± 2.0% vs 3.13% vs 0.9% respectively） （p 〈0.01） , and the comparison between group A and B showed no statistical difference. It is concluded that the CXCR4 specific siRNA can effectively down-regulate the CXCR4 mRNA expression, which induces the cell apoptosis and cell cycle arrest, thereby inhibits the Jurkat cell proliferation.

关 键 词：CXCR4基因 JURKAT细胞 细胞凋亡 细胞周期 RNA干扰 

分 类 号：R733.7[医药卫生—肿瘤]