腺病毒介导反义IGF-1抑制鼠胶质瘤细胞增殖的实验研究

Adenovirus mediated antisense IGF-1 gene therapy for rat glioma

目的重组腺病毒反义ＩＧＦ－１基因（ＡｄＨＣＭＶ－ＩＧＦ－１Ａ）治疗大鼠Ｃ６胶质瘤。方法体外克隆反义ＩＧＦ－１３００ｂＰ基因片段重组入腺病毒，经扩增纯化，测定滴度，体外体内感染Ｃ６胶质瘤细胞，观察ＩＧＦ－１的表达及肿瘤大小、生存期变化。结果重组腺病毒反义ＩＧＦ－１滴度６．５×１０１０ｐｆｕ／ｍｌ，反义ＩＧＦ－１基因序列正确，体外Ｃ６－Ａｄ－ＩＧＦ－１Ａ的ＩＧＦ－１表达降低，Ａｄ－ＩＣＦ－１Ａ治疗皮下Ｃ６肿瘤细胞接种组，肿瘤完全消失。与对照组相比有显著性差异（Ｐ＜０．０１），Ａｄ－ＩＧＦ－１Ａ治疗脑内Ｃ６肿瘤细胞接种组，大鼠生存期超过９０天，而对照组分别为１６．５±１．４天（Ａｄ－ｌａｃｚ组）、１５．７±１．６天（生理盐水组），Ｐ＜０．００１。结论重组腺病毒反义ＩＧＦ－１可抑制Ｃ６细胞ＩＧＦ－１的表达，ＡｄＨＣＭＶ－ＩＧＦ－１Ａ治疗大鼠Ｃ６胶质瘤效果显著。

Objective To observe the treatment effect of rat C6 glioma with recombinant adenovirus antisense IGF-1 gene (AdHCWV-IGF-1A). Methods Antisense IGF-1 gene of 300 base pairs was coloned into adenovirus. The recombinant adenovirus was amplified, purified and assessed of the titer. The C6 glioma cells were transfected by the recombinant adenovirus in vitro and in vivo. The expression of IGF-1 in vitro and tumor size, survival oftumor-bearing rats were observed. Results The bier of recombinat adenovirus was 6.5 ×1010 pfu/ml. The expression of IGF-1 in C6 cells transfected by AdHCMV-IGF-1A was decreased. When the rats of C6 subcutaneons gliomawere treated with AdHCMV-IGF-1A, the tumors all disappeared while as the tumor size of control was significantlylarge. When the rats of C6 intracranial glioma were treated with AdHCMV-IGF-1A, AdlacZ, saline solution the survivals were 90 days, 16.5±1 .4 days, 15.7±1 .5 days respectively (P < 0.001). Conclusion The expressionof IGF-1 of C6 could be suppressed by AdHCMV-IGF-1A. Recombinant adenovirus antisense IGF-1 had great effect intreating at c6 glioma in vivo.