Guo Rui, Zhang M, Xi Yun, Ma Yufei, Liang Sheng, Shi Shuo, Miao Ying, Li Biao
Department of Nuclear Medicine, Rui Jin Hospital, School of medicine, Shanghai JiaoTong University, Shanghai, China.
Department of Nuclear Medicine, Xin Hua Hospital, School of medicine, Shanghai JiaoTong University, Shanghai, China.
PLoS One. 2014 Jul 7;9(7):e102011. doi: 10.1371/journal.pone.0102011. eCollection 2014.
To investigate the role of 188Re in human sodium iodide symporter (hNIS) theranostic gene-mediated human glioma imaging and therapy in model mice.
The human glioma cell line U87 was transfected with recombinant lentivirus encoding the hNIS gene under the control of cytomegalovirus promoter (U87-hNIS). The uptake and efflux of 188Re were determined after incubating the cells with 188Re. 188Re uptake experiments in the presence of various concentrations of sodium perchlorate were carried out. In vitro cell killing tests with 188Re were performed. U87-hNIS mediated 188Re distribution, imaging and therapy in nude mice were also tested.
U87-hNIS cell line was successfully established. The uptake of 188Re in U87-hNIS cells increased up to 26-fold compared to control cells, but was released rapidly with a half-life of approximately 4 minutes. Sodium perchlorate reduced hNIS-mediated 188Re uptake to levels of control cell lines. U87-hNIS cells were selectively killed following exposure to 188Re, with a survival of 21.4%, while control cells had a survival of 92.1%. Unlike in vitro studies, U87-hNIS tumor showed a markedly increased 188Re retention even 48 hours after 188Re injection. In the therapy study, there was a significant difference in tumor size between U87-hNIS mice (317±67 mm3) and control mice (861±153 mm3) treated with 188Re for 4 weeks (P<0.01).
The results indicate that inserting the hNIS gene into U87 cells is sufficient to induce specific 188Re uptake, which has a cell killing effect both in vitro and in vivo. Moreover, our study, based on the function of hNIS as a theranostic gene allowing noninvasive imaging of hNIS expression by 188Re scintigraphy, provides detailed characterization of in vivo vector biodistribution and level, localization, essential prerequisites for precise planning and monitoring of clinical gene therapy that aims to individualize gene therapy concept.
探讨188Re在人钠碘同向转运体（hNIS）治疗诊断基因介导的模型小鼠人胶质瘤成像及治疗中的作用。
用编码在巨细胞病毒启动子控制下的hNIS基因的重组慢病毒转染人胶质瘤细胞系U87（U87-hNIS）。用188Re孵育细胞后，测定188Re的摄取和流出。进行了在不同浓度高氯酸钠存在下的188Re摄取实验。用188Re进行体外细胞杀伤试验。还测试了U87-hNIS介导的188Re在裸鼠体内的分布、成像及治疗。
成功建立了U87-hNIS细胞系。与对照细胞相比，U87-hNIS细胞中188Re的摄取增加了26倍，但释放迅速，半衰期约为4分钟。高氯酸钠将hNIS介导的188Re摄取降低至对照细胞系水平。暴露于188Re后，U87-hNIS细胞被选择性杀伤，存活率为21.4%，而对照细胞存活率为92.1%。与体外研究不同，即使在注射188Re后48小时，U87-hNIS肿瘤中188Re的滞留也显著增加。在治疗研究中，用188Re治疗4周后，U87-hNIS小鼠（317±67 mm3）和对照小鼠（861±153 mm3）的肿瘤大小有显著差异（P<0.01）。
结果表明，将hNIS基因插入U87细胞足以诱导特异性188Re摄取，其在体外和体内均具有细胞杀伤作用。此外，我们基于hNIS作为治疗诊断基因的功能，通过188Re闪烁显像对hNIS表达进行无创成像的研究，提供了体内载体生物分布和水平、定位的详细特征，这是精确规划和监测旨在实现基因治疗个体化概念的临床基因治疗的必要前提。
