利用DNA克隆和重组技术,构建增殖腺病毒SG600-IL24。ELISA检测比较SG600-IL24和非增殖腺病毒Ad-IL24分别感染肝癌细胞BEL-7402、Hep3B、HepGⅡ、SK-Hep-1和成纤维细胞BJ细胞后IL24的表达量,表明SG600-IL24病毒能在肿瘤细胞内大量表达IL-24,最高可达773 ng/mL(MOI=1),且在多数肿瘤细胞内IL24表达量显著高于Ad-IL24,而在正常细胞内表达量与Ad-IL24基本相同。半数组织感染剂量(TCID50)法检测SG600-IL24 48 h和96 h的增殖情况,发现SG600-IL24可在上述4种肿瘤细胞内大量增殖,96 h的最高增殖倍数可达到246153。MTT法和CPE法对比SG600-IL24和Ad-IL24对不同细胞的杀伤作用结果显示上述4种肿瘤细胞SG600-IL24对上述4种肿瘤细胞的50%杀伤剂量(IC50)和90%杀伤剂量(IC90)均明显低于Ad-IL24。以上结果表明SG600-IL24体现了病毒治疗与基因治疗的双重抗肿瘤用,其效果明显好于Ad-IL24,为该病毒治疗肝癌的体内研究打下基础。
SG600-IL24 is constructed by DNA cloning and recombination techniques. Then ELISA assay is performed to measure IL-24 gene expression in different liver cancer cell lines BEL-7402, Hep3B, HepG Ⅱ , SK-Hep-1, normal cell line BJ. The expression of IL24 in four tumor cells produced by SG600- IL24 is larger than that by Ad-IL24, and the highest concentration of IL24 generated by SG600-IL24 can reach 773 ng/mL, while the two virus have the basically same effect on normal cells. Viral replleation in different cell lines is evaluated by TCID50. After being infected for 48h and 96h, SG600-IL24 repelieates largely in the four different cancer cells, and the largest number of replicative folds is 246153 after 96h. In vitro anti-tumor effects of SG600-IL24 and Ad-IL24 to liver cancer cell lines are analyzed by MTT and CPE. The results demonstrate that SG600-IL24 has higher tumor killing effect on different tumor cells than Ad-IL24, which suggests great significance of SG600-IL24 in potent anti-tumor application associated with non-specific killing and SG600-IL-24 has great utility as a therapeutic agent for different types of cancers in clinic in future.