摘要
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: Two recombinant bacu loviruses expressing green fluorescent protein (GFP) and hNIS, respectively, were constructed according to the Bac-to-Bac Baculovirus Expression System package. The transduction efficiency of human hypopharyngeal cancer cells (FaDu cells) by Bac-GTP was determined by flow eytometry, and the loealizaton of hNIS on FaDu cells was determined by indirect im- munofluorescence. The iodine uptake, dynamic iodine uptake and inhibition of iodine uptake by NaCIO were examined. The radioactivity and fluorescence intensity in Bac-GFP and Bac-hNIS co-infected FaDu cells were measured. Results: Bae-GFP and Bac-hNIS were constructed successfully. The transduction efficiency of FaDu cells by Bac-GFP at MOI (multiplicity of infection) 400 with sodium butyrate could reach 95.16%. NIS protein could be localized accurately on FaDu cell membranes and performing its function of iodine uptake. The correlation coefficient between radioactivity and fluorescence intensity was 0.943, a linear positive correlation, and the change in level of radioactivity could be used to es- timate the change in fluorescence intensity. Conclusions: Baculovirus could efficiently transduce FaDu cells. Iodine up- take by FaDu cells could be mediated effectively by Bae-hNIS, and NIS could be used for imaging and diagnosis of human hypopharyngeal squamous cell carcinoma.
出处
《诊断学理论与实践》
2013年第4期426-431,共6页
Journal of Diagnostics Concepts & Practice
基金
2011年上海市科委基金(114119a6400)
关键词
钠碘转运体
杆状病毒
人下咽鳞癌
显像
Sodium/iodine symporter
Baculovirus
Human hypopharyngeal squamous cell carcinoma
Imaging