Sodium butyrate enhances the expression of baculovirus-mediated sodium/iodide symporter gene in A549 lung adenocarcinoma cells
Sodium butyrate enhances the expression of baculovirus-mediated sodium/iodide symporter gene in A549 lung adenocarcinoma cells
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丁酸钠增强A549肺腺癌细胞中杆状病毒介导的钠/碘同向转运蛋白基因的表达
DOI：
10.1097/mnm.0b013e32833dedd7
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发表时间：
2010-10-01
影响因子：
1.5
通讯作者：
Li, Biao
中科院分区：
文献类型：
--
作者：
Guo, Rui;Zhang, Yifan;Li, Biao
目的钠/碘同向转运体（NIS）表达的增加是报告基因成像和有效的放射性碘治疗肿瘤所必需的。我们研究了丁酸钠是否可以通过新开发的杆状病毒转移人NIS（hNIS）基因在A549人肺腺癌中诱导碘积累增加。方法构建含hNIS基因的重组杆状病毒[Bac-cytomegalovirus（CMV）-hNIS]，该基因受CMV启动子控制。将Bac-CMV-hNIS转染A549细胞后，用免疫荧光和western blot检测hNIS蛋白的表达。在存在或不存在丁酸钠的情况下，用125 I-碘化物孵育转染的细胞后，测定碘的摄取和流出。结果经丁酸钠处理后，转染Bac-CMV-hNIS的A549细胞hNIS蛋白表达增加。Bac-CMV-hNIS转染的A549细胞比未转染的细胞积累了约9倍的125 I，丁酸钠以剂量依赖方式增加125 I摄取量（P<0.001）。然而，观察到放射性的快速流出，在含125 I的培养基被非放射性培养基取代后的前2分钟内损失了50%。结论丁酸钠可提高杆状病毒介导的hNIS报告基因肺腺癌显像的效率。然而，需要定义额外的条件，以减少放射性核素治疗目的的放射性碘的快速流出。
ObjectiveIncreased expression of sodium/iodide symporter (NIS) is required for reporter gene imaging and effective radioiodine treatment of tumor. We investigated whether increased accumulation of iodine can be induced by sodium butyrate through a newly developed baculoviral transfer of the human NIS (hNIS) gene in A549 human lung adenocarcinoma. MethodsA recombinant baculovirus [Bac–cytomegalovirus (CMV)–hNIS] encoding hNIS gene under the control of the CMV promoter was constructed. After A549 cells were transfected with Bac–CMV–hNIS in the presence of sodium butyrate, the expression of hNIS protein was detected by immunofluorescence and western blot analysis. The uptake and efflux of iodine were determined after the incubation of the transfected cells with 125I-iodide in the presence or absence of sodium butyrate. ResultsImmunocytochemical staining and western blot analysis showed increased hNIS protein expression in A549 cells transfected with Bac–CMV–hNIS after sodium butyrate treatment. Bac–CMV–hNIS transfected A549 cells accumulated up to about nine times more 125I than nontransfected cells; the amount of 125I uptake increased in a sodium butyrate in dose-dependent manner (P<0.001). However, rapid efflux of radioactivity was observed, with 50% lost during the first 2 min after 125I-containing medium had been replaced by a nonradioactive medium. ConclusionOur results indicated that an improved efficiency of baculovirus-mediated hNIS reporter gene imaging in lung adenocarcinoma is possible with treatment with sodium butyrate. However, additional conditions need to be defined to reduce the rapid efflux of radioiodine for the purpose of radionuclide therapy.