hNIS转基因介导^(131)碘对鼻咽癌细胞增殖和凋亡的影响  

Effect of ^(131)I on proliferation and apoptosis of nasopharygeal carcinoma cell transferred human sodium/iodide symporter gene

作　　者：钟兴[1] 弓健[2] 曾祥凤[3] 徐浩[2] 

机构地区：[1]暨南大学附属第一医院医学影像中心,广东广州510630 [2]暨南大学附属第一医院核医学科,广东广州510630 [3]暨南大学组织与免疫移植实验中心,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2010年第4期387-392,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省重大科技计划项目(2003A3080501)

摘　　要：目的:克隆人的甲状腺钠/碘同向转运体(hNIS)基因,研究其转导在鼻咽癌细胞内的功能,以及131碘对鼻咽癌细胞增殖和凋亡的影响。方法:利用逆转录和聚合酶链反应(RT-PCR)从毒性弥漫性甲状腺肿(Graves病)病人的甲状腺组织中扩增得到hNIS的可编码区基因,并克隆到pcDNA3.1(+)-FLAG载体,获得重组真核表达质粒pcDNA3.1(+)-hNIS-FLAG。采用脂质体转染的方法,将pcDNA3.1(+)-hNIS-FLAG质粒导入到鼻咽癌细胞株CNE2,G418筛选得到稳定表达hNIS鼻咽癌细胞系CNE2-hNIS,体外培养条件下检测其对放射性碘的摄取和外流情况。CCK8试剂盒检测131碘对鼻咽癌细胞增殖的作用,AnnexinⅤ-FITC/PI双标记流式细胞技术(FCM)检测131碘作用后鼻咽癌细胞凋亡情况。结果:成功克隆hNIS基因,并建立能稳定表达hNIS的鼻咽癌细胞系CNE2-hNIS。CNE2-hNIS对125碘的吸收量比CNE2高约15倍,但在无碘的环境中,细胞内滞留的碘外流迅速,有效半衰期约8min。131碘在72 h后对CNE2-hNIS的增殖产生抑制作用,细胞早期凋亡率增加,并随着131碘浓度的增加,作用逐渐增强(P<0.01)。结论:从Graves病人的甲状腺组织中克隆的hNIS基因转染鼻咽癌细胞后,能使鼻咽癌细胞发挥高水平吸碘功能,131碘能够抑制转染hNIS的鼻咽癌细胞的增殖,促进细胞凋亡。Aim：To clone sodium/iodide symporter（hNIS） gene of human thyroid tissues and to investigate its function in nasopharygeal carcinoma cell and effect of 131I on proliferation and apoptosis of nasopharygeal carcinoma cells. Methods： The sodium/iodide symporter gene was amplified from human thyroid tissues of the patients with Grave＇s disease by RT-PCR,and cloned into pcDNA 3.1（＋）-FLAG plasmid.The recombinate eukaryotic expression plasmid of pcDNA 3.1（＋）-hNIS-FLAG was reconstructed.The pcDNA 3.1（＋）-hNIS-FLAG was transduced into CNE2 by lipofectamine 2000.After screening by G418,stable transfected cell line was established.The uptake and efflux of iodide was examined in vitro.The proliferation rates of cells was detected by CCK8 and the apoptosis rates of cells was detected by flow cytometry with annexinⅤ-FITC and PI double labeling.Results： The cloned hNIS cDNA sequence was identical to the published sequence.The stable transfected cell line CNE2-hNIS was established.The absorbed doses of 125I in CNE2-hNIS were 15 times higher than in CNE2.The efflux of iodide was also rapid（T1/2eff=8 min）.CCK8 showed 131I inhibited the growth of CNE2-hNIS at 72 h and the inhibition effect was dose-dependent（P0.01）.AnnexinV-FITC/PI assay showed that CNE2-hNIS cells apoptosis after 72 h by 131I（P0.01）.Conclusion：After the nasopharygeal carcinoma cells are transfected by cloned hNIS gene sourced from thyroid tissues of the patient with Grave＇s disease,the transfected tumor cells can induce a high function level to absorb iodum.131I can inhibit proliferation and enhance apoptosis of the nasopharygeal carcinoma cells.

关 键 词：人钠/碘同向转运体 基因转染 碘放射性同位素 

分 类 号：R736.1[医药卫生—肿瘤]