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目的探讨下调CXCR4的表达对U87胶质瘤细胞周期和凋亡的影响,为CXCR4 sh RNA治疗胶质瘤提供实验依据。方法设计合成CXCR4的特异性sh RNA,采用脂质体转染U87细胞株。实验将细胞分为空白对照组(A组),阴性对照组(转染非特异载体p GPU6/GFP/NC组)(B组),实验组(转染CXCR4-sh RNA组)(C组)共三组,用RT-PCR检测各组细胞CXCR4基因表达情况,用流式细胞术检测细胞周期分布和细胞的凋亡情况。结果成功构建了CXCR4的特异性sh RNA。C组的U87细胞CXCR4 m RNA为(42.5±4.6)%,显著低于B组的(65.4±5.9)%和A组的(69.6±7.4)%(P<0.05),G0/G1期细胞比例增加(39.4±1.7)%与(25.8±1.3)%和(20.3±1.2)%,G2/M期细胞比例相应下降(20.2±1.6)%与(30.1±1.2)%和(26.4±1.3)%;S期细胞比例相应下降(46.5±2.7)%与(55.7±2.8)%和(54.9±2.6)%,凋亡细胞率为(18.68±0.44)%明显高于A组(5.14±0.23)%和B组(4.87±0.16)%(P<0.05)。结论 CXCR4 sh RNA能够有效下调U87细胞CXCR4基因表达,诱导细胞凋亡和细胞周期阻滞,从而抑制细胞的增殖,提示CXCR4在胶质瘤发展过程中具有重要作用。
Objective To investigate the effects of CXCR4 expression on the cell cycle and apoptosis in U87 glioma cells and to provide experimental evidence for the treatment of gliomas by CXCR4 sh RNA. Methods The specific sh RNA of CXCR4 was designed and synthesized and transfected into U87 cells by liposome. The cells were divided into blank control group (group A), negative control group (transfected with nonspecific vector p GPU6 / GFP / NC group) (group B), experimental group (transfected with CXCR4-sh RNA group) A total of three groups were detected by RT-PCR CXCR4 gene expression in each group, using flow cytometry to detect cell cycle distribution and cell apoptosis. Results The specific sh RNA of CXCR4 was successfully constructed. The CXCR4 mRNA of U87 cells in group C was (42.5 ± 4.6)%, which was significantly lower than that in group B (65.4 ± 5.9%) and group A (69.6 ± 7.4)% (P <0.05) (20.2 ± 1.6)% and (30.1 ± 1.2)% and (26.4 ± 1.3)% of the cells in G2 / M phase were increased by (39.4 ± 1.7)% and (25.8 ± 1.3)% and (20.3 ± 1.2) (46.5 ± 2.7)% and (55.7 ± 2.8)% and (54.9 ± 2.6)%, respectively. The rate of apoptotic cells was (18.68 ± 0.44)% higher than that of group A (5.14 ± 0.23)% And group B (4.87 ± 0.16)% (P <0.05). Conclusion CXCR4 sh RNA can effectively down-regulate CXCR4 gene expression in U87 cells, induce apoptosis and cell cycle arrest, and thus inhibit cell proliferation, suggesting that CXCR4 plays an important role in glioma development.