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目的研究慢病毒载体携带的血管内皮细胞生长因子受体2(VEGFR2)基因短发夹RNA(Lenti6/sh VEGFR2)对VEGFR2表达的特异抑制及对HL60人白血病细胞所诱导血管生成的作用。方法2005年10月至2006年2月,中山大学附属第二医院儿科选择6周龄、体重25~30g的雄性NOD/SCID小鼠15只,分成3组,实验采用双盲法。将pU6/sh VEGFR2入门克隆与pLenti6/BLOCK-iTTM-DEST载体发生LR重组反应。用HL60人白血病细胞评价Lenti6/sh VEGFR2对血管生成的作用。通过检测骨微血管密度评估Lenti6/sh VEGFR2感染HL60细胞后对VEGFR2活性的特异抑制作用。结果建立了表达VEGFR2基因小发夹RNA的慢病毒载体(Lenti6/sh VEGFR2)。在静脉异体移植的HL60白血病小鼠中,Lenti6/sh VEGFR2感染显著抑制了血管生成(P<0.05)。结论慢病毒介导的VEGFR2 RNA干扰有可能成为通过抑制病理性血管生成治疗白血病的有效方法。
Objective To study the specific inhibition of VEGFR2 expression by lentiviral vector carrying Lenti6 / sh VEGFR2 gene and its effect on angiogenesis induced by HL60 human leukemia cells. Methods From October 2005 to February 2006, 15 male NOD / SCID mice, 6 weeks old and 25-30 g, were selected from the Second Affiliated Hospital of Sun Yat-sen University and divided into three groups. The experiment was double-blind. The pU6 / sh VEGFR2 entry clone was subjected to an LR recombination reaction with the pLenti6 / BLOCK-iTTM-DEST vector. The effect of Lenti6 / sh VEGFR2 on angiogenesis was evaluated using HL60 human leukemia cells. The specific inhibitory effect of Lenti6 / sh VEGFR2 on the activity of VEGFR2 was evaluated by detecting the microvessel density. Results Lentiviral vector (Lenti6 / sh VEGFR2) expressing small hairpin RNA of VEGFR2 gene was constructed. Lenti6 / sh VEGFR2 infection significantly inhibited angiogenesis (P <0.05) in HL60 leukemic mice that were xenotransplanted. Conclusion Lentivirus-mediated VEGFR2 RNA interference may be an effective method for the treatment of leukemia by inhibiting pathological angiogenesis.