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目的:探讨杆状病毒介导的人钠碘转运体(human sodium/iodine symporter,hNIS)应用于人下咽鳞癌显像和诊断的可行性。方法:通过Bac-to-Bac杆状病毒表达系统包装分别含GFP和hNIS的重组杆状病毒(Bac-GFP和Bac-hNIS),用流式细胞仪检测Bac-GFP对人下咽癌细胞(FaDu)的转导效率,并用间接免疫荧光检测hNIS在FaDu细胞中的定位。进行hNIS介导的碘摄取、动态摄碘和NaClO4碘摄取抑制实验,并检测共转染了Bac-GFP和Bac-hNIS的FaDu细胞中的荧光强度和放射性计数。结果:在丁酸钠的作用下,Bac-GFP在感染复数为400时,对FaDu细胞的转导效率可达95.16%;hNIS可准确定位于FaDu细胞膜上,具备摄碘功能;放射性计数与荧光强度间呈线性正相关(相关系数为0.943),可通过放射性计数的变化来反映荧光强度的变化。结论:杆状病毒对FaDu细胞具有高转导效率,Bac-hNIS可有效介导FaDu细胞的碘摄取,hNIS可用于人下咽鳞癌的显像和诊断。
Objective: To investigate the feasibility of using baculovirus-mediated human sodium / iodine symporter (hNIS) for imaging and diagnosis of human hypopharyngeal squamous cell carcinoma. Methods: Recombinant baculovirus (Bac-GFP and Bac-hNIS) containing GFP and hNIS were packaged by Bac-to-Bac baculovirus expression system. The expression of Bac-GFP FaDu) transduction efficiency, and indirect immunofluorescence assay hNIS localization in FaDu cells. The hNIS-mediated iodine uptake, dynamic iodine uptake and NaClO4 iodine uptake inhibition assays were performed and the fluorescence intensity and radioactivity counts were measured in FaDu cells co-transfected with Bac-GFP and Bac-hNIS. Results: Under the action of sodium butyrate, the transfection efficiency of Bac-GFP was up to 95.16% in FaDu cells at a multiplicity of infection of 400. hNIS could be accurately localized on FaDu cell membrane with iodine-uptake function. The radioactivity count and fluorescence There was a linear positive correlation between intensities (correlation coefficient of 0.943), and the change of fluorescence intensity could be reflected by the change of radioactive count. CONCLUSION: Baculovirus has high transduction efficiency on FaDu cells. Bac-hNIS can effectively mediate iodine uptake in FaDu cells. HNIS can be used in the imaging and diagnosis of human hypopharyngeal squamous cell carcinoma.