CAJ | 학술논문

目的构建携带IL-24基因的溶瘤腺病毒CNHK600-IL24,评估该病毒在裸鼠体内对乳腺癌的抑瘤能力.方法将IL-24基因插入腺病毒穿梭载体SG502-△CR2,与5型腺病毒骨架载体pPE3共转染至293细胞,获得溶瘤腺病毒CNHK600-IL24.建立乳腺癌原位成瘤模型、尾静脉注射及左心室注射模拟转移瘤模型,通过尾静脉注射CNHK600-IL24,应用"活体内光学成像系统"动态地观察病毒的疗效.结果 CNHK600-IL24经测序及PCR鉴定正确,滴度为1.9×1010pfu/ml.乳腺癌原位成瘤模型:对照组的光子数以及肿瘤体积均明显高于CNHK600-IL24各治疗组(P＜0.05).CNHK600-IL24治疗后肿瘤组织出现明显的坏死,肿瘤细胞发生显著的凋亡,免疫组化检测可见肿瘤细胞内Hexon和IL-24的表达.尾静脉注射模拟转移瘤模型:对照组的裸鼠大部分于38 d前死亡,而CNHK600-IL24组的生存天数明显延长(P＜0.05).左心室注射模拟转移瘤模型:活体内光学成像可见对照组与治疗组之间具有明显差别.结论高滴度的溶瘤腺病毒CNHK600-IL24对于乳腺癌具有明显的抑瘤效果.
목적 구건휴대IL-24기인적용류선병독CNHK600-IL24,평고해병독재라서체내대유선암적억류능력.방법 장IL-24기인삽입선병독천사재체SG502-△CR2,여5형선병독골가재체pPE3공전염지293세포,획득용류선병독CNHK600-IL24.건립유선암원위성류모형、미정맥주사급좌심실주사모의전이류모형,통과미정맥주사CNHK600-IL24,응용"활체내광학성상계통"동태지관찰병독적료효.결과 CNHK600-IL24경측서급PCR감정정학,적도위1.9×1010pfu/ml.유선암원위성류모형:대조조적광자수이급종류체적균명현고우CNHK600-IL24각치료조(P＜0.05).CNHK600-IL24치료후종류조직출현명현적배사,종류세포발생현저적조망,면역조화검측가견종류세포내Hexon화IL-24적표체.미정맥주사모의전이류모형:대조조적라서대부분우38 d전사망,이CNHK600-IL24조적생존천수명현연장(P＜0.05).좌심실주사모의전이류모형:활체내광학성상가견대조조여치료조지간구유명현차별.결론 고적도적용류선병독CNHK600-IL24대우유선암구유명현적억류효과.
Objective To construct an oncolytic adenovirus CNHK600-IL24, and to observe the in vivo effects of CNHK600-IL24 in treating breast cancer. Methods The IL-24 gene was cloned into adenovirus shuttle vector SG502-△CR2, and CNHK600-IL24 was obtained by cotransfection of SG502-INSIL24 and pPE3 plasmids and subsequent recombination in 293 cells. Based on the establishment of the athymic mice model of breast cancer in situ and imitated metastatic breast cancer by injection in the vena caudalis and the left artrium, we administered the virus by the tail vein. We used the optical imaging in vivo system to monitor the effects. Results The oncolytic adenovirus CNHK600-IL24 was correctly constructed and confirmed by restriction DNA sequence analysis and PCR. The titer of CNHK600-IL24 reached 1.9 ×1010pfu/ml. Establishing athymic mice model of breast cancer in situ, the volume and photon number of the tumors in the control group was significantly larger than those of the CNHK600-IL24 group( P ＜0. 05). The tumor had conspicuous necrosis after the treatment of CNHK600-IL24. There was noticeable apoptosis of the tumor cells. Immunohistochemistry showed the expression of IL-24 and the Hexon protein in the tumor cells.In athymic mice model of imitated metastatic breast cancer by infusion into the vena caudalis, most of the mice in the control group died before 38 days, the mice of the CNHK600-IL24 group survived significantly longer(P ＜0. 05 ). Using athymic mice model of imitated metastatic breast cancer by infusion in the left artrium, the optical imaging in vivo system showed obvious difference between the control group and the CNHK600-IL24 group. Conclusions The high-titer oncolytic adenovirus CNHK600-IL24 was successfully constructed and purified. The oncolytic adenovirus had obvious antitumor effect on breast cancer.