人IL-2重组腺病毒转染人肺腺癌细胞的体外生物学特性及体内抗肿瘤研究

The biological characteristics in vitro and anti-tumor effect in vivo of lung cancer cells infected with recombinant adenovirus expressing hIL-2

目的 :研究感染hIL 2重组病毒的人肺腺癌细胞 (Anip973)的体外生物学特性及小鼠体内抗肿瘤作用。方法 :将携带有hIL 2基因的重组腺病毒 (rAd hIL 2 )感染人肺腺癌细胞系 ,通过细胞生长实验、克隆形成实验等观察其对Anip973肿瘤细胞的作用 ;利用PCR技术对转染后的细胞进行检测并应用ELISA试剂盒检测转染后的细胞IL 2的分泌量 ;通过肿瘤局部注射rAd hIL 2的方法观察其在小鼠体内的抗肿瘤作用。结果 :rAd hIL 2经扩增、纯化后 ,滴度可达 10 1 0 PFU ml,当病毒量为30MOI时 ,对Anip973细胞的转染率达 90 %以上。转染的Anip973肿瘤细胞其生长能力、克隆形成率等无明显变化。 2 4小时细胞培养上清IL 2的分泌量为 2 0pg 1ml 2× 10 5细胞。体内实验表明注射rAd hIL 2的小鼠肿瘤生长缓慢 ,体积明显小于对照组 (P <0 0 5 ) ,生存期显著延长。结论 :腺病毒介导的细胞因子基因hIL 2转染的Anip973肿瘤细胞体外生物学特性未见明显变化 ,而体内实验则有明显的抗肿瘤作用。

Objective:To study the biological characteristics of tumor cells infected with recombinant adenovirus expressing hIL 2, and investigate the antitumor effect of the rAd Methods:Human lung adenocarcinoma cell line Anip973 was infected with recombinant adenovirus expressing hIL 2(rAd hIL 2) Cell growth assay, colony formation were used to observe the effects on tumor cells hIL 2 gene was transduced into cancer cells mediated by recombinant adenovirus, then detected by PCR and agarose gel electrophoresis and its products were detected by ELISA assay The intratumor injection of rAd LacZ, rAd hIL 2 was carried out to evaluate its antitumor effects Results:The titer of rAd reached 10 10 PFU/ml and more than 90% Anip973 cells could be infected by 30MOI rAd Growth inhibition assay, colony formation assay showed no significant difference compared with that of the control cells The 24 h IL 2 expression increased greatly Antitumor study indicated that the tumor grew slowly in the bearing tumor mice treated with rAd Tumor volume was significantly smaller and survive time was prolonged than that of controls Conclusion:There were no significantly changes of tumor cells infected with recombinant adenovirus expressing hIL 2 The intratumor injection of rAd LacZ?rAd hIL 2 could inhibit the growth of solid tumor