Department of Nuclear Medicine, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, No. 197, Rui Jin 2nd Road, Shanghai 200025, China.
Technol Cancer Res Treat. 2014 Apr;13(2):139-48. doi: 10.7785/tcrt.2012.500368. Epub 2013 Aug 2.
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 5 400). 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 125I uptake amount in HepG2 cells after co-vector administration with Bac-NIS and Bac-GFP at different MOIs, and found a high correlation coefficient (r(2) = 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.
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较差。本研究旨在探讨杆状病毒载体将目的基因转导到肝癌细胞中的效率，并评估通过共载体给药方法使用杆状病毒载体递送钠碘同向转运体 (NIS) 基因作为报告基因来监测肝癌基因治疗中靶治疗基因表达的可行性。我们构建了（绿色荧光蛋白）GFP 和 NIS 表达的杆状病毒载体（Bac-GFP 和 Bac-NIS），并测量了 HepG2 细胞和其他肿瘤细胞（A549、SW1116 和 8505C）中杆状病毒的转导效率，结果表明，随着病毒感染复数（MOI）的增加，转导效率和靶基因表达水平在 HepG2 细胞中升高，HepG2 细胞的转导效率明显高于其他肿瘤细胞（MOI = 200 时为 60.8%）。此外，在较高的 MOI（MOI 5 400）下，杆状病毒转导对 HepG2 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 HepG2 细胞表现出高效的碘摄取，可被高氯酸钠（NaClO₄）抑制。并且我们测量了 Bac-NIS 和 Bac-GFP 在不同 MOI 下共载体给药后 HepG2 细胞的荧光强度和 125I 摄取量的相关性，发现相关性系数很高（r² = 0.8447），为成功评估杆状病毒介导的 NIS 报告基因监测肝癌治疗中靶基因表达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研究肝癌细胞中传递治疗基因的潜在载体。并且使用杆状病毒载体递送 NIS 基因作为报告基因来监测靶基因的表达是可行的。因此，它为未来杆状病毒介导的肝癌细胞治疗基因传递或治疗基因表达监测提供了一种有效的方法和良好的基础。
