内皮抑素重组腺病毒治疗肝癌裸鼠模型的实验观察

Experimental Research on the Treatment of Adenovirus-mediated Human Endostatin for Liver Cancer in Nude Mice

目的研究观察内皮抑素重组腺病毒Ad-endostatin对BALB/c裸鼠接种HepG2肝癌细胞肿瘤模型的治疗作用。方法用重组腺病毒Ad-endostatin感染HUVEC细胞,用MTT法和Western Blot法分别分析HUVEC细胞的增殖抑制作用和Endostatin蛋白表达。用Ad-endostatin接种HepG2肝癌细胞裸鼠模型,观察小鼠的肿瘤大小变化,用免疫组化法观察肿瘤组织中血管内皮细胞CD31的表达。结果用Ad-endostatin 50感染HU-VEC细胞24、48h后,HUVEC细胞活性分别下降到(79.30±3.76)%、(63.75±2.82)%,差异具有统计学意义(P<0.05)。Ad-Endo接种HepG2肝癌细胞裸鼠21d后,肿瘤体积为435±26.67mm3,体积抑制率达51.40%,差异显著(P<0.001);Ad-Endo治疗组肿瘤的CD31阳性细胞数68.34±3.29,差异显著(P<0.001)。结论Ad-endostatin能明显抑制肝癌细胞生长,明显抑制肿瘤组织中血管内皮细胞生成,对小鼠荷瘤局部具有明显治疗作用。

Objective To study the therapeutic effect of adenovirus-mediated human endostatin on Liver Cancer in BALB/e nude mice bearing HepG2 cells. Methods Cell viability and the expression of endostatin in HUVEC were analyzed by MTI＂ and Western blot, respectively. BALB/c nude mice bearing HepG2 cells were treated by Ad-endostatin,Ad-EGFP and PBS, once every 3 days. The tumor volume and the body weight were observed. CD31 in microvascular endothelial cells of tumor tissue was analyzed by immunohistochemistry. Re- sults The viability of HUVEC cells was （79.30±3.76） %, （ 63.75-2.82 ） %, respectively, after infected by 50 MOI Ad-endostatin for 24,48h. The proliferation of HUVEC cells was obviously inhibited by Ad-endostatin, compared with 10 MOI Ad-endostatin（P 〈0.05）. The tumor volume was （435±26.67）mm3 ,after 30 days of HepG2 cells inoculation,the tumor growth inhibition rate was 51.40%. The positive cells of CD31 in tumors was （68.34±3.29） in Ad-Endo group, the tumor angiogenesis and the mean vessel density index was obvious- ly decreased in the established tumor, compared with control group and Ad-EGFP group（ P 〈 0.001 ）. Conclu- sion Ad-endostatin can inhibit the tumor growth of BALB/c nude mice, and decrease obviously angiogenesis in tumor. Adenovirns-mediated human endostation has some therapeutic effect on nude tumor mice bearing liver cancer cells,