山东医药2017，Vol.57Issue(26)：9-12,4.DOI:10.3969/j.issn.1002-266X.2017.26.003
以脐带间充质干细胞为载体的靶向基因治疗系统联合5-FU对裸鼠HepG2移植瘤的作用
Effect of targeted gene therapeutic system with HUMSCs as vectors combined with 5-FU on HepG2 exnograft tumor of nude mice
摘要
Abstract
Objective To construct a targeted gene therapy system with human umbilical cord mesenchymal stem cells (HUMSCs) as vectors, and to observe its therapeutic effect on nude mice HepG2 xenografts combined with low-dose 5-fluorouracil (5-FU).Methods HUMSCs were isolated from fresh sterile neonatal umbilical cord by tissue adherent method.We used the methods of PCR, overlap PCR, and restriction enzyme cleavage and linkage to construct the lentiviral expression vector pLentiR.E1A and the adenovirus shuttle vector pAd-hTERTp-IL24.The migratory ability to HepG2 cells of HUMSCs co-infected by lentivirus and adenovirus in vitro was determined by Transwell experiment.HepG2 transplanted tumor model was established in BALB/c nude mice by subcutaneous inoculation.The tumor-bearing mice were divided into five groups and the groups were separately injected with PBS, HUMSCs, low-dose 5-FU, HUMSCs infected by lentivirus combined with ow-dose 5-FU, and HUMSCs co-infected by double virus combined with 5-FU.The anti-tumor effect of each group was compared.The apoptosis of hepatocellular carcinoma cells induced by adenovirus Ad-hTERTp-IL24 and low-dose 5-FU was detected by BD Annexin-Ⅴ-PE apoptotic kit.Results The lentiviral expression vector pLentiR.E1A and adenovirus expression vector pAd-hTERTp-IL24 were successfully constructed and the lentivirus LentiR.E1A and adenovirus Ad-hTERTp-IL24 were successfully packaged.Transwell experiment showed that there was no significant change in the migratory ability of HUMSCs to tumor cells after co-infected by double virus.In treatment of HepG2 exnograft tumor of nude mice, the tumor volume was significantly smaller in the treatment group than in the control group (P<0.01).Ad-hTERTp-IL24 combined with low-dose 5-FU induced tumor apoptosis in the tumor microenvironment, and its fluorescence intensity was stronger than that of the other treatment control groups.Conclusion The targeted gene therapeutic system based on HUMSCs is successfully established, and adenovirus Ad-hTERTp-IL24 combined with low-dose 5-FU have significant inhibitory effect on HepG2 exnograft tumor of nude mice.关键词
脐带间充质干细胞/腺病毒/靶向基因治疗系统/5-氟尿嘧啶/HepG2移植瘤Key words
human umbilical cord mesenchymal stem cells/adenovirus/targeted gene therapeutic system/5-fluorouracil/HepG2 exnograft tumor分类
医药卫生引用本文复制引用
孔凡妮,查剑英,杨圆圆,卢杨,李真真,张砚君..以脐带间充质干细胞为载体的靶向基因治疗系统联合5-FU对裸鼠HepG2移植瘤的作用[J].山东医药,2017,57(26):9-12,4.基金项目
国家科技重大专项基金资助项目(2012ZX09102301-015). （2012ZX09102301-015）
