Potential Usefulness of Baculovirus-Mediated Sodium-Iodide Symporter Reporter Gene as Non-Invasively Gene Therapy Monitoring in Liver Cancer Cells: An In Vitro Evaluation
Potential Usefulness of Baculovirus-Mediated Sodium-Iodide Symporter Reporter Gene as Non-Invasively Gene Therapy Monitoring in Liver Cancer Cells: An In Vitro Evaluation
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DOI：
10.7785/tcrt.2012.500368
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发表时间：
2014-04
影响因子：
2.8
通讯作者：
Yu Pan;Hai-fei Wu;Shuai Liu;Xiaoping Zhou;Hong-yan Yin;Biao Li;Yifan Zhang
中科院分区：
文献类型：
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作者：
Yu Pan;Hai-fei Wu;Shuai Liu;Xiaoping Zhou;Hong-yan Yin;Biao Li;Yifan Zhang
原发性肝癌是所有癌症中死亡率最高的癌症之一，目前的主要治疗方法预后不良。本研究旨在检测杆状病毒载体将目的基因导入肝癌细胞的效率，并探讨杆状病毒载体通过共载体给药的方式将钠碘转运体（NIS）基因作为报告基因用于肝癌基因治疗的可行性。我们构建（绿色荧光蛋白）表达GFP和NIS的杆状病毒载体（Bac-GFP和Bac-NIS），并测量HepG 2细胞和其它肿瘤细胞中的杆状病毒转导效率（A549、SW 1116和8505 C），并且其显示在HepG 2细胞中的转导效率和靶基因表达水平随着病毒感染复数（MOI）的增加而升高，HepG 2细胞的转导效率（MOI = 200时为60.8%）显著高于其他肿瘤细胞。此外，杆状病毒转导在较高MOI（MOI = 400）下对HepG 2细胞没有细胞毒性。我们还进行了动态碘摄取试验，发现Bac-NIS转导的HepG 2细胞表现出有效的碘摄取，可以被高氯酸钠（NaClO 4）抑制。并测定了不同MOI下共转染Bac-NIS和Bac-GFP后HepG 2细胞荧光强度与125 I摄取量的相关性，相关系数为0.8447，为成功评价杆状病毒介导的NIS报告基因监测靶基因表达在肝癌治疗中的可行性提供了良好的基础。因此，本研究表明杆状病毒载体是一种潜在的载体，用于研究肝癌细胞的治疗基因。利用杆状病毒载体携带NIS基因作为报告基因，对目的基因的表达进行监测是可行的。这为杆状病毒介导的治疗性基因传递或治疗性基因表达监测在肝癌细胞中的应用研究提供了一种有效的方法和良好的基础。
Primary liver cancer has one of the highest mortality rates of all cancers, and the main current treatments have a poor prognosis. This study aims to examine the efficiency of baculovirus vectors for transducing target gene into liver cancer cells and to evaluate the feasibility of using baculovirus vectors to deliver the sodium-iodide symporter (NIS) gene as a reporter gene through co-vector administration approach to monitor the expression of the target therapeutic gene in liver cancer gene therapy. We constructed (green fluorescent protein) GFP- and NIS-expressing baculovirus vectors (Bac-GFP and Bac-NIS), and measured the baculovirus transduction efficiency in HepG2 cells and other tumor cells (A549, SW1116 and 8505C), and it showed that the transduction efficiency and target gene expression level rose with increasing viral multiplicity of infection (MOI) in HepG2 cells, and HepG2 cells had a significantly higher transduction efficiency (60.8% at MOI = 200) than other tumor cells. Moreover, the baculovirus transduction was not cytotoxic to HepG2 cells at a higher MOI (MOI = 400). We also performed dynamic iodide uptake trials, and found that Bac-NIS-transduced HepG2 cells exhibited efficient iodide uptake which could be inhibited by sodium perchlorate (NaClO4). And we measured the correlation of fluorescent intensities and 125 I uptake amount in HepG2 cells after co-vector administration with Bac-NIS and Bac-GFP at different MOIs, and found a high correlation coefficient (r2 = 0.8447), which provides a good basis for successfully evaluating the feasibility of baculovirus-mediated NIS reporter gene monitoring target gene expression in liver cancer therapy. Therefore, this study indicates that baculovirus vector is a potential vehicle for delivering therapeutic genes in studying liver cancer cells. And it is feasible to use a baculovirus vector to deliver NIS gene as a reporter gene to monitor the expression of target genes. It therefore provides an effective approach and a good basis for future baculovirus-mediated therapeutic gene delivering or therapeutic gene expression monitoring in liver cancer cells studies.