Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, No. 197, Rui Jin 2nd Road, Shanghai 200025, China.
World J Gastroenterol. 2010 Nov 14;16(42):5367-74. doi: 10.3748/wjg.v16.i42.5367.
To investigate the feasibility of radionuclide therapy of colon tumor cells by baculovirus vector-mediated transfer of the sodium/iodide symporter (NIS) gene.
A recombinant baculovirus plasmid carrying the NIS gene was constructed, and the viruses (Bac-NIS) were prepared using the Bac-to-Bac system. The infection efficiency in the colon cancer cell line SW1116 of a green fluorescent protein (GFP) expressing baculovirus (Bac-GFP) at different multiplicities of infection (MOI) with various concentrations of sodium butyrate was determined by flow cytometry. An in vitro cytotoxicity assay was also conducted after infection of SW1116 cells with Bac-NIS. Iodine uptake of Bac-NIS infected SW1116 cells and inhibition of this uptake by sodium perchlorate was examined, and the effect of Bac-NIS-mediated (131)I in killing tumor cells was evaluated by cell colony formation tests.
Infection and transgene expression in SW1116 with Bac-GFP were significantly enhanced by sodium butyrate, as up to 72% of SW1116 cells were infected with the virus at MOI of 400 and sodium butyrate at 0.5 mmol/L. No obvious cytotoxicity was observed under these conditions. Infection of SW1116 with Bac-NIS allowed uptake of (131)I in these tumor cells, which could be inhibited by sodium perchlorate. The viability of SW1116 cells infected with Bac-NIS was significantly lower than with Bac-GFP, suggesting that NIS gene-mediated (131)I uptake could specifically kill tumor cells.
Baculovirus vector-mediated NIS gene therapy is a potential approach for treatment of colon cancer.
研究杆状病毒载体介导的钠/碘同向转运体（NIS）基因转染对结肠肿瘤细胞放射性核素治疗的可行性。
构建携带 NIS 基因的重组杆状病毒质粒，利用 Bac-to-Bac 系统制备病毒（Bac-NIS）。用不同感染复数（MOI）和不同浓度丁酸钠的绿色荧光蛋白（GFP）表达杆状病毒（Bac-GFP）感染结肠癌细胞系 SW1116，用流式细胞术检测其感染效率。用 Bac-NIS 感染 SW1116 细胞后进行体外细胞毒性试验。检测 Bac-NIS 感染的 SW1116 细胞摄取碘的情况以及高氯酸钠对此摄取的抑制作用，并用细胞集落形成试验评价 Bac-NIS 介导的（131）I 对杀伤肿瘤细胞的作用。
SW1116 细胞用丁酸钠感染和转染基因表达显著增强，MOI 为 400 时，高达 72%的 SW1116 细胞被感染，丁酸钠浓度为 0.5mmol/L。在这些条件下未观察到明显的细胞毒性。SW1116 细胞用 Bac-NIS 感染后，这些肿瘤细胞可以摄取（131）I，高氯酸钠可以抑制其摄取。与 Bac-GFP 相比，Bac-NIS 感染的 SW1116 细胞活力明显降低，表明 NIS 基因介导的（131）I 摄取可以特异性杀伤肿瘤细胞。
杆状病毒载体介导的 NIS 基因治疗是治疗结肠癌的一种有潜力的方法。
