人TNF-α重组腺病毒转染人肺腺癌细胞的体外生物学特性研究(英文)

The in-vitro biological characteristics of lung cancer cells infected with recombinant adenovirus expressing hTNF-α

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INF α重组病毒的人肺腺癌细胞 (Anip973)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 hTNF α经扩增、纯化后 ,滴度可达 10 1 0PFU 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。

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 α transduced into cancer cells mediated by recombinant adenovirus were detected by PCR and agarose gel electrophoresis. Results The titers of rAd reached to 10 10 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 significantly changes of tumor cells infected with recombinant adenovirus expressing hTNF α.