携带EGFP报告系统的五型腺病毒对肝癌干细胞的感染效率

Study on the Efficiency of 5-type Adenovirus Which Carrying a EGFP Report System Infection with Liver Stem Cells

利用携带EGFP报告系统的五型腺病毒(Ad5-EGFP)作为工具,以发生上皮-间质转化(epithelial-mes-enchymal transition,EMT)前后的肝癌细胞系Huh7为研究对象,采用流式细胞仪、荧光定量PCR等技术开展腺病毒对发生EMT前后的肝癌细胞系Huh7的感染效率的对比研究。实验结果表明:Ad5-EGFP对上皮生产因子β1(TGF-β1)处理前的Huh7细胞系感染率比TGF-β1处理后的细胞系感染率更高。同时Ad5-EGFP的主要受体-CAR在肝癌细胞系Huh7发生EMT后表达有所降低。证实了肿瘤干细胞更难被腺病毒侵染,为了适应临床治疗的需要,腺病毒作为基因治疗的载体需要进一步被改造。

The 5-type adenovirus （Ad5-EGFP） infection efficiency in Hepatoma cell line Huh7 between before and after happening EMT is studied by Flow cytometry and qRT-PCR. The results show that the infection of Ad5-EGFP in normal Huh7 cells is higher than that in Huh7 cells after treated with TGF-β1. In addition, CAR receptor, which is the primary receptor of Ad5, is expressed in lower level after EMT happening HuhT. It suggests that cell stem cell is harder to be infected by adenovirus, and adenovirus should be improved better in order to be applied in clinical treatment.