本文在利用COS/TPC同源重组法高效制备人CD80重组腺病毒的基础上 ,对感染人CD80重组腺病毒的肺癌细胞的体外生物学特性进行研究。方法 :以缺陷型重组腺病毒为载体 ,将人CD80基因导入肺癌细胞 ,利用PCR和琼脂糖电泳对肺癌细胞中人CD80基因进行了检测 ,以FACS、电镜等方法观察分析肺癌细胞被rAd CD80感染前后的变化。结果 :人CD80重组腺病毒经扩增、纯化后 ,滴度可达 10 10 PFU/ml ,当病毒为 30MOI时 ,腺病毒对肺癌细胞Anip973的转染率达 90 %以上。转染rAdCD80的肺癌细胞除表面结构和超微结构有轻微变化外 ,其生长能力及克隆形成率无明显变化 ,rAdCD80对肺癌细胞的增殖周期亦无影响。结论 :以上结果为制备肿瘤疫苗打下基础。

Objective: To investigate in vitro the biological characteristics of AdCD80-infected human lung cancer cells on the basis of generation of replication-deficient hB7-1(CD80) recombinant adenovirus. Methods: Human CD80 gene was transduced into lung cancer cells mediated by recombinant adenovirus and then the expression of the gene was detected by PCR and agarose gel electrophoresis. The biological characteristics of the above cells were analysed with electron microscope, FACS and etc. Results: The titers of rAd reached to 10 10 PFU/ml and more than 90% Anip973 lung cancer cells could be infected by 30 MOI rAd. The growth curve and cloning efficiency of rAdCD80-infected Anip973 cells showed no significant difference compared with that of the control cells. The cell proliferation cycle of rAdCD80-infected 973 cells showed no change through FACS test. Having been infected by rAdCD80, the surface structure and ultrastructure of 973 cells had a little change. Conclusion: These results will lay foundation for tumor vaccines.

R735.7

本课题受国家“九五”重点（96-906-10-20）攻关计划资助及黑龙江省自然科学基金（D9723）资助