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目的观察腺病毒介导的人血管抑素基因对胰腺癌的治疗作用。方法通过病毒重组技术将人血管抑素基因克隆入增殖缺陷型腺病毒基因组中,获得腺病毒滴度达5．5×10~(10) pfu/ ml,观察转染表达后的生物学活性,通过建立裸鼠动物模型(每组数n=15例),分析基因转导后胰腺癌组织中血管抑素的表达情况及对肿瘤血管的抑制作用。结果构建了血管抑素的重组腺病毒载体pCA13-hAG；检测到血管抑素在体外mRNA水平和蛋白质水平表达率分别为87%和81%,得到279 bp电泳条带和38 000大小的蛋白条带；荷瘤裸鼠体内肿瘤体积显著低于对照组(P＜0．05),治疗组MVD为9．85±1．20,两对照组肿瘤微血管密度(MVD)分别为20．35±2．15、17,66±2．34 (P＜0．05)。结论所构建的pCA13-hAG重组腺病毒载体可有效表达具有生物学活性的血管抑素,使肿瘤内微血管生成减少,肿瘤细胞增殖减慢,为抗血管生成治疗实体瘤的临床应用奠定基础。
Objective To observe the therapeutic effect of adenovirus-mediated human angiostatin gene on pancreatic cancer. Methods Human angiostatin gene was cloned into the genome of proliferating defective adenovirus by virus recombination technique. The titer of adenovirus was 5.5 × 10 ~ (10) pfu / ml. The biological activity after transfection was observed. The animal model of nude mice (n = 15 in each group) was established to analyze the expression of angiostatin and its inhibitory effect on tumor vessels after gene transduction. Results The angiostatin recombinant adenovirus vector pCA13-hAG was constructed. The expression of angiostatin at mRNA and protein levels in vitro was 87% and 81%, respectively. A 279 bp electrophoresis band and a 38,000-sized protein band (P <0.05). The MVD in the treatment group was 9.85 ± 1.20, and the MVD in the two control groups was 20.35 ± 2 .15,17,66 ± 2.34 (P <0.05). CONCLUSION: The constructed pCA13-hAG recombinant adenovirus vector can effectively express the biologically active angiostatin, reduce the intra-tumor angiogenesis and slow down the proliferation of tumor cells, and lay a foundation for the clinical application of anti-angiogenesis therapy for solid tumors.