摘要
目的研究腺相关病毒转染人内皮抑素基因对肝癌细胞体内致瘤作用的影响。方法用含人内皮抑素基因的重组腺相关病毒(rAAV2/ES)体外转染人肝癌细胞Hep3B,以空病毒(AAV2)转染作阴性对照,RT-PCR方法检测ES在mRNA水平的表达。将转染细胞接种裸鼠,通过ELISA方法检测裸鼠血液中人内皮抑素的的浓度。观察内皮抑素对移植瘤的生长抑制作用。结果RT-PCR结果显示腺相关病毒可介导内皮抑素基因高效转染Hep3B细胞;rAAV2/ES转染细胞接种裸鼠后内皮抑素的表达随时间逐渐升高,在第4周达到最高浓度(115·79±8·70)ng/ml;rAAV2/ES转染组8只裸鼠中6只成瘤,成瘤时间为(26·80±6·42)d,明显长于对照组(17·17±4·17)d(P<0·05);对裸鼠移植瘤检测显示瘤体重量和瘤内微血管密度(MVD)分别为(0·36±0·22)g、(3·50±1·05)个/视野,明显低于对照组(1·54±0·48)g、(12·67±1·97)个/视野(P<0·01)。结论腺相关病毒介导人内皮抑素基因转染可有效抑制Hep3B细胞在裸鼠体内的生长。
Objective To explore the effects of adeno-associated virus-mediated gene transfer of human endostatin on carcinogenesis of hepatocellular carcinoma （HCC） cells in nude mice. Methods Hep3B cells were injected by recombinant adeno-associated virus with endostatin gene （rAAV2/ES） and empty virus （AAV2） was used to serve as the control. RT-PCR was employed to detect the expression of human endostatin gene at mRNA level. The transfected Hep3B cells were inoculated into the nude mice, then the expression of human endostatin was determined ELISA. The inhibitory effects of endostatin on the growth of xenograft were assessed. Results RT-PCR confirmed that rAAV2/ES efficiently infected Hep3B cells. After inoculation with transfected Hep3B cells, the endostatln expression level increased gradually with time prolongation and reached its peak （115.79±8.70 ng/ml） in the 4th week. In rAAV2/ES-transfected group, 6 out of the 8 mice had tumor and the time of tumor appearance was markedly longer than that in the control group （26.80±6.42 d vs. 17.17±4.17 d, P d0.05）. The tumor weight and microvessel density （MVD） were 0.36 ± 0.22 g and 3.50± 1.05/ HP, respectively. They were significantly lower than those in the control group （P〈0.01）. Conclusions Adeno-associated virus-mediated gene transfer of human endostatin can effectively inhibit the erowth of HeD3B cell xenograft in nude mice.
出处
《中华肝胆外科杂志》
CAS
CSCD
2005年第12期840-843,共4页
Chinese Journal of Hepatobiliary Surgery
关键词
癌
肝细胞
内皮抑素
基因
腺相关病毒
Carcinoma, hepatocellular
Endostatin
Gene
Adeno-associated virus
Nude mouse
