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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Bel-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。
Objective To investigate the inhibitory effect of Ad-MMP2AS carrying antisense type 2 matrix metalloproteinase (MMP2) gene on human hepatocellular carcinoma animal model and angiogenesis. Methods Human hepatocellular carcinoma cell line (Bel-7402) was infected with Ad-MMP2AS. Boyden Chamber was used to detect the inhibitory effect of Ad-MMP2AS on the degradation of Matrigel in Bel-7402 cells; Western blotting and gelatin zymography were used to determine the effect of Ad-MMP2AS on the secretion of MMP2 by Bel-7402 cells; -7402 cells were inoculated subcutaneously in nude mice to observe its tumorigenicity; Ad-MMP2AS intratumoral injection to observe its inhibitory effect on the growth of hepatocellular carcinoma; Tissue sections, HE staining to observe the growth of tumor cells; Immunohistochemical analysis of tumor tissue vascular density . Results Ad-MMP2AS-infected Bel-7402 cells through the Matrigel Bel-7402 cells decreased by 52%, 4.3 times decreased tumor formation; intratumoral injection of Ad-MMP2AS tumor growth was reduced by 63%; tumor tissue density decreased by 2.6 Times Conclusion Ad-MMP2AS can inhibit the growth of hepatocellular carcinoma and tumor angiogenesis, and has the therapeutic potential for hepatocellular carcinoma.