NIS基因转染对肝细胞癌NIS蛋白表达及摄碘功能的影响  

Effects of NIS gene transfection on NIS protein expression and iodine uptake in hepatocellular carcinoma

作　　者：范义湘 易紫薇 胡煜麟 张宏嘉 林美珍 陈洁芳 肖汉[2] FAN Yixiang;YI Ziwei;HU Yulin;ZHANG Hongjia;LIN Meizhen;CHEN Jiefang;XIAO Han(Department of Nuclear Medicine,the Fifth Affiliated Hospital of Southern Medical University,Guangzhou 510900,China;Department of Nuclear Medicine,Guangdong Second Provincial General Hospital,Guangzhou 510317,China)

机构地区：[1]南方医科大学第五附属医院核医学科,广东广州510900 [2]广东省第二人民医院核医学科,广东广州510317

出　　处：《中国医学物理学杂志》2021年第10期1219-1222,共4页Chinese Journal of Medical Physics

摘　　要：目的:探讨肝细胞癌细胞核素靶向治疗方法。方法:提取NIS基因片段并构建重组质粒pcDNA3/NIS,将重组质粒导入肝细胞癌HepG2细胞。转染后24 h利用Western-Blot法检测HepG2细胞NIS蛋白表达,采用^(125)I结合试验评估转染后细胞的摄碘率,采用DAPI染色法评估HepG2细胞摄^(125)I后的凋亡情况。实验组与对照组之间NIS蛋白表达、摄碘率以及细胞凋亡率比较采用t检验。结果:蛋白电泳表明经NIS基因转染后,实验组HepG2细胞已表达NIS蛋白,表达强度显著高于对照组(t=2.693,P<0.05)。实验组HepG2细胞摄碘率B/T%平均为(18.4±5.8)%,显著高于对照组(t=36.842,P<0.05)。结合^(125)I后24 h,实验组凋亡细胞数多于对照组,平均凋亡率为(19.2±5.3)%,显著高于对照组(t=3.086,P<0.05)。结论:转染外源性NIS基因可上调肝细胞癌HepG2细胞NIS蛋白表达,使其具备摄碘功能,加快细胞凋亡,为放射性碘靶向治疗提供实验依据。Objective To explore the method of targeted radionuclide therapy for hepatocellular carcinoma.Methods The NIS gene fragment was extracted,and the recombinant plasmid pCDNA3/NIS was constructed.The recombinant plasmid was introduced into hepatocellular carcinoma HepG2 cells.The NIS protein expression in HepG2 cells at 24 h after transfection was detected by Western-Blot assay;and the iodine uptake rate of the transfected cells was evaluated by ^(125)I binding assay;and the apoptosis of HepG2 cells after ^(125)I uptake was evaluated by DAPI staining.NIS protein expression,iodine uptake rate and apoptosis rate were compared between experimental group and control group using t test.Results Protein electrophoresis showed that after NIS gene transfection,NIS protein was expressed in HepG2 cells in experimental group,and the expression intensity was significantly higher than that in control group(t=2.693,P<0.05).The iodine uptake rate(B/T%)of HepG2 cells in experimental group was(18.4±5.8)%,significantly higher than that of control group(t=36.842,P<0.05).At 24 h after ^(125)I uptake,the number of apoptotic cells in experimental group was larger than that in control group,and the average apoptosis rate in experimental group was(19.2±5.3)%,significantly higher than that in control group(t=3.086,P<0.05).Conclusion The transfection of exogenous NIS gene can up-regulate NIS protein expression in hepatocellular carcinoma HepG2 cells,enable the cells to have the function of iodine uptake,and accelerate cell apoptosis.The study provide an experimental basis for targeted radionuclide therapy.

关 键 词：肝细胞癌 钠碘转运体 放射性碘治疗 NIS基因 摄碘功能 细胞凋亡 

分 类 号：R34[医药卫生—基础医学] R817.1