摘要
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 In the present study, a lentiviral vector expressing small hairpin RNA against the VEGFR2 gene （Lenti6/sh VEGFR2） was constructed to specifically inhibit VEGFR2 expression and to evaluate the effect of Lenti6/sh VEGFR2 on angiogenesis of HL60 human leukemia cell. Methods Recombine pU6/sh VEGFR2 entry clone with Lenti6/BLOCK-iTTM-DEST vector using LR reaction. The effect of Lenti6/sh VEGFR2 on angiogenesis was evaluated in an HL60 human leukemia cell line. Lenti6/sh VEGFR2 infection of HL60 cells specifically inhibited the VEGFR2 activity as determined by evaluation of the microvessel density of the bone. Results A lentiviral vector expressing small hairpin RNA against the VEGFR2 gene （ Lenti6/sh VEGFR2 ） was constructed, In an HL60 intravenous xenograft leukemia mouse model, Lenti6/sh VEGFR2 infection dramatically suppressed angiogenesis （ P 〈 0. 05 ）. Conclusion Lentiviralmediated VEGFR2 RNA interference potentially represents an effective therapeutic strategy against leukemia by inhibiting pathological angiogenesis.
出处
《中国实用儿科杂志》
CSCD
北大核心
2008年第2期106-108,I0001,共4页
Chinese Journal of Practical Pediatrics
关键词
血管生成
人血管内皮细胞
白血病
SHRNA
血管内皮细胞生长因子受体
Angiogenesis
Human vascular endothelial cell
Leukemia
Small hairpin RNA
Vascular endothelial growth factor receptor
