携突变减毒Stx1编码序列重组腺病毒的构建及其抗乳腺癌的活性  

Construction of recombinant replication defective adenoviral vectors carrying attenuated Shiga-like toxin Ⅰ mutant gene and their therapeutic effects on breast cancer

作　　者：安秀梅[1] 魏枫[1] 于津浦[1] 李慧[1] 杨莉莉[1] 任秀宝[1] 

机构地区：[1]天津医科大学附属肿瘤医院免疫学研究室,乳腺癌防治教育部重点实验室,天津市"肿瘤防治"重点实验室,天津300060

出　　处：《中国肿瘤生物治疗杂志》2009年第3期258-262,共5页Chinese Journal of Cancer Biotherapy

基　　金：天津市卫生局科技基金(No.05KY32);天津医科大学附属肿瘤医院博士启动基金(No.07B02)~~

摘　　要：目的:构建携带1/100、1/1000减毒活性的突变减毒志贺样毒素Ⅰ(Shiga-like toxin1,Stx1)编码序列的复制缺陷型腺病毒,并观察其抗人乳腺癌细胞裸鼠移植瘤的活性。方法:重叠PCR法构建毒性为原毒素毒性1/100、1/1000的突变减毒Stx1编码序列,T-A克隆并测序后构建携带该编码序列的复制缺陷型腺病毒载体Adv-Stx-1R170L。制备人乳腺癌T47D细胞移植瘤裸鼠模型,Adv-Stx-1R170L肿瘤局部注射给药,评价其体内抑瘤能力。结果:经测序证实正确克隆了1/100、1/1000减毒活性的突变减毒Stx1编码序列,成功构建携带该突变减毒Stx1编码序列的复制缺陷型腺病毒载体Adv-Stx-1R170L。体内实验显示,Adv-Stx-1R170L可以有效抑制裸鼠体内移植瘤的生长,与携带绿色荧光蛋白编码序列的腺病毒对照组、PBS对照组相比,差异有统计学意义(P<0.05)。结论:成功构建了携带1/1000原毒素活性的突变减毒Stx1编码序列的重组复制缺陷型腺病毒载体,该病毒载体可以有效抑制裸鼠体内移植瘤的生长,且未见明显毒性作用。Objective：To construct recombinant replication defective adenoviral vectors encoding 1/100 and 1/1000 attenuated Sbiga-like toxin Ⅰ （Stxl） gene, and to study their therapeutic effects against breast cancer T47D cells in vivo. Methods： Genes encoding 1/100 or 1/1000 attenuated Stxl were amplified by overlapping PCR and were cloned into T vectors. The inserted gene was verified by nucleotide sequencing. Replication defective adenoviral vector Adv-Stx-I R170L containing 1/1000 attenuated Stxl mutant gene was generated by AdMAX Adenoviral Vector System. Nude mouse models bearing human breast cancer T47D cells were established, and the tumor inhibitory effect of Adv-Stx-1 R170L was studied by intra-tumor injection of the recombinant adenovirus. Results： Vectors carrying 1/100 or 1/1000 attenuated Stx-1 gene were successfully constructed and were verified by nucleotide sequencing. Recombinant replication defective adenoviral vector Adv-Stx-1RI70L containing 1/1000 attenuated Shiga-like toxin I gene was constructed. In vivo study showed that AdvStx-1Rl70L significantly inhibited the growth of implanted T47D tumor in the nude mice compared with Adv-GFP and PBS group（P 〈 0.05 ）. Conclusion ： Recombinant replication defective adenoviral vector Adv-Stx-1 R170L encoding 1/1000 attenuated Shiga-like toxin I gene has been successfully constructed, and it can effectively inhibit the growth of implanted T47D tumor in nude mice, without obvious toxicity.

关 键 词：志贺样毒素Ⅰ 突变减毒 乳腺癌 复制缺陷型腺病毒 

分 类 号：R737.9[医药卫生—肿瘤] R730.54[医药卫生—临床医学]