腺病毒介导反义基质金属蛋白酶-2抑制肝癌生长和血管生成的研究

目的探讨携带反义二型基质金属蛋白酶（MMP2）基因的重组腺病毒（Ad-MMP2AS）对人肝癌动物模型生长和血管生成的抑制作用。方法用我们构建的Ad-MMP2AS感染人肝癌细胞株（Bel-7402）。Boyden Chamber检测Ad-MMP2AS对Bd-7402细胞降解人工基底膜（Matrigel）的抑制作用；Western blotting和明胶酶谱分析测定Ad-MMP2AS对Bel-7402细胞分泌MMP2的影响；用感染Ad-MMP2AS的Bel-7402细胞接种于裸鼠皮下观察其成瘤能力；Ad-MMP2AS瘤内注射。观察它对肝癌生长的抑制作用；肿瘤组织切片、HE染色观察瘤细胞生长情况；免疫组织化学分析肿瘤组织血管密度。结果Ad-MMP2AS感染的Bel-7402细胞穿过Matrigel的Bel-7402细胞数下降52％、成瘤量下降4.3倍；瘤内注射Ad-MMP2AS使瘤体生长减少63％；肿瘤组织血管密度减少2.6倍。结论Ad-MMP2AS能抑制肝癌的生长和肿瘤血管的生成，对肝癌有治疗潜力。

Inhibitory effect of antisense matrix metalloproteinase-2 mediated by adenovirus on hepatocellular carcinoma growth and angiogenesis in vivo

Objective To investigate the inhibitory effect of a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2(MMP2) on the growth and angiogenesis of human hepatocellular carcinoma (HCC) animal model in vivo.Methods The recombinant adenoviral vector carrying antisense MMP2(Ad-MMP2AS) which was constructed previously was used to infect the human HCC cell line(Bel-7402).The invasiveness of the Bel-7402 cells was assayed in Matrigel,and the production of MMP2 in the Bel7402 cells was detected by using Western blotting analysis and gelatin zymography.Following the Ad-MMP2AS-infected cells were subcutaneously inoculated in nude mice,and injected intratumorally into pre-existing tumors,the tumors were removed,sectioned,stained with H&E and stained with an anti-CD31 monoclonal antibody.Results Compared with PBS or Ad-CMV-infected cells,infection of Bel-7402 cells with Ad-MMP2AS significantly reduced MMP2 enzyme activity,the number of Bel-7402 cells penetrating Matrigel was decreased by 52%,and the tumor volume was reduced by 4.3-fold.Direct intratumoral injection of Ad-MMP2AS into pre-existing tumors could decrease the tumor growth by 63% and the tumor vessel density was reduced by 2.6-fold.Conclusion The recombinant adenovirus carrying antisense MMP2 can effectively inhibit the invasiveness and growth of Bel-7402 cells in vitro and in vivo.