靶向肝癌HepG2细胞VEGF基因siRNA表达载体裸鼠成瘤抑制作用  

Inhibitory effect of HepG2 VEGF-siRNA on the growth of human liver cancer cell xenograft in nude mice

作　　者：黄秋林[1] 刘璇[1] 阳勇[2] 曹超[1] 戴小明[1] 韩东[1] 雷俊悦[1] 

机构地区：[1]南华大学附属第一医院普通外科,湖南衡阳421001 [2]南华大学附属第二医院普通外科,湖南衡阳421001

出　　处：《中国现代医学杂志》2013年第14期18-22,共5页China Journal of Modern Medicine

基　　金：湖南省自然科学基金(No:11JJ3114);湖南省科技计划立项项目(No:2010SK3034)

摘　　要：目的探讨靶向肝癌HepG2细胞VEGF基因siRNA表达载体对人肝癌细胞株裸鼠成瘤的抑制作用。方法将设计并合成构建好的靶向肝癌HepG2细胞VEGF基因的psuper.retro.neo-VEGF-siRNA表达载体转入HepG2细胞,通过G418抗性筛选出稳定株(HepG2/psuper.retro.neo-VEGF-siRNA,实验组),同时设对照组(HepG2/psuper.retro.neo组)和空白组(HepG2组)。分别移植BALA/c裸鼠成瘤,计算各组鼠成瘤潜伏期和接种20 d后瘤重,Western-blotting检测肿瘤组织中VEGF的表达变化,免疫组织化学SP法检测瘤组织内微血管密度(MVD)。结果实验组、对照组和空白组裸鼠成瘤潜伏期分别为(9.2±1.2)、(3.9±0.7)和(3.8±0.9)d;接种20 d后3组平均瘤重分别为(194±57)、(566±86)和(626±96)g;瘤组织VEGF蛋白表达水平分别为(0.075±0.012)、(0.198±0.009)和(0.205±0.008);MVD分别为(13.6±2.8)、(34.3±2.9)和(35.3±3.5),实验组与对照组和空白组比较差异有统计学意义(P<0.05)。结论合成和构建的靶向人肝癌HepG2 VEGF基因干扰质粒具有抑制人肝癌细胞裸鼠成瘤和血管生成的作用。【Objective】 To investigate the inhibitory effect of small interfering RNA（siRNA） targeting HepG2 vascular endothelial growth factor（VEGF） on the growth of human liver cancer cell xenograft in nude mice.【Methods】 VEGF siRNA targeting HepG2 expression vector constructed was transfected into HepG2 cells（the expression vector of VEGF siRNA targeting HepG2 which has been constructed was transfected into HepG2 cells） as experimental group（HepG2/psuper.retro.neo-VEGF-siRNA cells）;psuper.retro.neo was transfected into HepG2 cells as control group,and untransfected HepG2 cells as empty group.Stable cell clones were screened by G418,and transplanted into nude mice to establish cancer xenograft.Tumor growth was monitored（monitoring the growth of tumor）.The expression of VEGF protein in tumor tissues was detected by Western-blot.Microvessel density（MVD） was detected by SP immunohistochemistry.【Results】 The time of tumor formation was longer in experimental group than in control group and empty group（9.2 ± 1.2 d,3.9 ± 0.7 d and 3.8 ± 0.9 d,P〈 0.05）.The weight of the xenograft tumor after 20 d transplanted was lighter in experimental group（after transplanted 20 days,the weight of the xenograft tumor in experimental group was lighter）than in control group and empty group（194 ± 57 g,566 ± 86 g and 626 ± 96 g,P〈 0.05）.The expression of VEGF protein and MVD were less in experimental group（in experimental group were less） than in control group and empty group（0.075 ± 0.012,0.198 ± 0.009,0.205 ± 0.008,and 13.600 ± 2.800,34.300 ± 2.900,35.300 ± 3.500,P〈 0.05）.【Conclusion】 VEGF siRNA expression vector constructed can effectively inhibit the growth of cancer xenograft and angiogenesis in nude mice.

关 键 词：肝癌 RNA干扰 血管内皮生长因子 微血管密度 

分 类 号：R735.7[医药卫生—肿瘤]