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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。
The recombinant adenovirus SG600-IL24 was constructed by DNA cloning and recombination techniques. The results of ELISA showed that the expression of IL24 in SG600-IL24 and non-proliferating adenovirus Ad-IL24 infected BEL-7402, Hep3B, HepGⅡ, SK-Hep-1 and fibroblast BJ cells, IL-24 was highly expressed in tumor cells up to 773 ng / mL (MOI = 1), and the expression of IL24 in most tumor cells was significantly higher than that in Ad-IL24. However, the expression level of IL-24 in normal cells was similar to that of Ad-IL24 the same. The proliferation of SG600-IL24 at 48 h and 96 h was detected by TCID50 method. It was found that SG600-IL24 could proliferate in the above four kinds of tumor cells with the highest multiplication of 246153 at 96 h. The killing effect of SG600-IL24 and Ad-IL24 on different cells by MTT assay and CPE assay showed that the killing rates of IC50 and 90% of the above four kinds of tumor cells by SG600-IL24 ) Were significantly lower than Ad-IL24. These results show that SG600-IL24 virus treatment and gene therapy with dual anti-tumor effect, the effect was significantly better than Ad-IL24, the virus for the treatment of liver cancer in vivo to lay the foundation.