【作者】 李天天； 李殿俊； 田长富； 刘旭；

【Author】 LI Tian-tian,LI Dian-jun,TIAN Chang-fu, LIU Xu (Department of Immunology, Harbin Medical University, Harbin 150086, China)

【机构】 哈尔滨医科大学免疫学教研室；

【摘要】 目的在利用COS／TPC同源重组法高效制备hTNF-α重组腺病毒基础上,对感染人INI-α组病毒的人肺腺癌细胞(Anip9r73)的体外生物学特性进行研究。方法将携带有人TNF-α基因的重组腺病毒(rAd-hTNF-α)感染人肺腺癌细胞系,通过细胞生长实验、克隆形成实验、流式细胞分析、形态学检查等观察其对Anip973肿瘤细胞的作用;利用PCR和琼脂糖电泳对肺癌中人TNF-α基因进行了检测。结果 rAd-hTNFα经扩增、纯化后,滴度可达1010 PFU／ml,当病毒为30MOI时,对Anip973细胞的转染率达90％以上。转染的Anip973肿瘤细胞除表面结构和超微结构有轻微变化外,其生长能力、克隆形成率及细胞周期等无明显变化。结论腺病毒介导的细胞因子基因hTNF-α转染的Anip973肿瘤细胞未见明显变化,为制备肿瘤疫苗打下基础。更多还原

【Abstract】 Objective To study the in-vitro biological characteristics of tumor cells infected with recombinant adenovirus expressing hTNF-α. Methods Human lung adenocarcinoma cell line Anip973 was infected with recombinant adenovirus expressing hTNF-α. Cell growth assay, colony formation, flowcytometry assay and morphology were used to observe the effects on tumor cells. HTNF-a transduced into cancer cells mediated by recombinant adenovirus were detected by PCR and agarose gel electrophoresis. Results The titers of rAd reached to 1010 PFU/ml and more than 90% Anip973 cells could be infected by 30MOI rAd. The surface structure and ultrastructure of tumor cells infected with rAd showed little change. The cell growth curve, cloning efficiency and the cell differential cycle of rhTNF-α-infected Anip973 cells showed no significant difference compared with that of the control cells. Conclusion There are no significandy changes of tumor cells infected with recombinant adenovirus expressing hTNF-α.更多还原