hNIS基因转导黑色素瘤细胞介导^(125)I摄取的实验研究  

Transfer of the human sodium/iodide symporter gene enhances iodide uptake in melanoma cells

作　　者：陈立波[1] 黄芳 费俭[2] 郭礼和[2] 陆汉魁[1] 朱瑞森[1] 

机构地区：[1]上海市第六人民医院核医学科,200233 [2]中国科学院上海细胞生物学研究所

出　　处：《中华核医学杂志》2003年第2期72-74,共3页Chinese Journal of Nuclear Medicine

摘　　要：目的　探讨克隆的人钠 碘同向转运体 (hNIS)基因cDNA的生物学性能及其用于黑色素瘤放射治疗的可能性。方法　用反转录 聚合酶链反应 (RT PCR)从人甲状腺组织总RNA中扩增出hNIS基因cDNA序列 ,将其克隆至pUCm T载体并亚克隆至真核载体pcDNA3中 ,电击法将重组质粒导入黑色素瘤细胞 (B16 )建立新细胞系 (B16 A) ,在体外培养条件下研究其对放射性碘的摄取及外流情况。结果　成功克隆了hNIS基因 ,建立了能稳定表达hNIS基因的新型细胞系B16 A和单转空质粒pcDNA3的细胞系B16 B。B16 A细胞的摄碘能力较B16细胞高 17倍 ,较单转空质粒pcDNA3细胞系B16 B高 19倍。碘的外流过程十分迅速 ,有效半衰期 (T1 2 )仅 10min。结论　体外培养条件下 ,hNIS基因转导黑色素瘤细胞其本身足以介导碘的摄取 ,但有效半衰期较短 ,有待进一步研究如何增加放射性碘的摄取量及延长其细胞内滞留时间 ,从而提高放射生物学效应。Objective To obtain human sodium/iodide symporter (hNIS) cDNA and to study its biological property and potential use as a therapeutic radioiodide for melanoma. Methods hNIS gene cDNA was amplified with total RNA from human thyroid tissue by RT PCR. The hNIS cDNA was inserted into cloning vector pUCm T and subcloned into eukaryotic expression vector pcDNA 3. The recombinant plasmid pcDNA 3 hNIS was introduced into B16 cells using the electroporation technique. The uptake and efflux of iodide was examined in vitro. Results The cloned hNIS cDNA sequence was identical to the published sequence. Two novel cell lines named B16 A containing hNIS and B16 B containing pcDNA 3 only were established. The resultant cell line B16 A accumulated 17 and 19 times more radioiodide in vitro than B16 and B16 B did, respectively. However the efflux of iodide from B16 A was also rapid ( T 1/2 =10 min). Conclusions Our preliminary data indicate that the transduction of the hNIS gene per se is sufficient to induce iodide transport in melanoma cells in vitro, but its T 1/2 is short. With regard to therapeutic application, however, further investigation is necessary so as to develop a method of maintaining more radioiodide in the cells for long enough to produce greater therapeutic effects.

关 键 词：hNIS基因 基因转导 黑色素瘤 细胞介导 摄取 碘放射性同位素 生物学性能 反转录-聚合酶链反应 

分 类 号：R817[医药卫生—影像医学与核医学]