^(125)I标记三链形成寡核苷酸对肝癌细胞生长的抑制作用  

Inhibition effects of ^(125)I-triplex forming oligonucleotide to hepatoma cells

作　　者：吕中伟[1] 侯敏[1] 何俊民[2] 蔡海东[1] 袁雪宇[1] 杨越华[1] 袁世栋[1] 

机构地区：[1]同济大学附属第十人民医院核医学科,上海200072 [2]同济大学生命科学与技术学院

出　　处：《中华核医学杂志》2007年第4期218-220,共3页Chinese Journal of Nuclear Medicine

基　　金：国家自然科学基金(30300092)

摘　　要：目的探讨^(125)I 标记的三链形成寡核苷酸(TFO)对肝癌细胞的抑制作用,为 HBV 相关肝细胞癌(HCC)的基因治疗提供实验依据。方法合成能与 HBV 前 S 基因特异结合的 TFO,并以^(125)I标记(^(125)I-TFO);分^(125)I-TFO、等量 TFO、相同活度^(125)I 和空白对照4组与 HepG2.2.15细胞共同培养,通过 ELISA 法检测培养前后各组细胞上清中乙型肝炎 e 抗原(HBeAg)和乙型肝炎表面抗原(HBsAg)含量的变化,以四甲基偶氮唑蓝(MTT)比色法检测各组细胞存活率。结果 ^(125)I-TFO 标记率>93%,稳定性较好,37%放置12 h 放化纯90.8%,48 h 81.1%,72 h 73.2%;^(125)I-TFO 组在转染48 h 对HepG2.2.15细胞合成 HBeAg 及 HBsAg 的抑制作用最强,对 HBeAg 和 HBsAg 表达的抑制率(74.5%和45.2%)及细胞杀伤率[72 h 细胞增殖抑制率为(31.6±2.5)%]高于其他实验组(P<0.01)。结论 ^(125)I-TFO 可以抑制 HBV 抗原表达,杀伤肝癌细胞,为制备抗 HBV、抗 HCC 的放射性基因治疗药物提供依据。Objective Triplex forming oligonucleotide （TFO） has been reported as a new antigene strategy. The purpose of this study was to observe the inhibition effects of ^125 I-TFO on hepatoma cells and to investigate the possibility of using ^125 I-TFO as an antigene radiotherapy technique for hepatocellular carcino ma （HCC） related to HBV. Methods TFO complementary to the initiator of S gene of HBV was synthesized and labeled with ^125 I. HepG2.2.15 cells, in which HBV genome was integrated, were incubated with ^125I-TFO, TFO and ^125I respectively. After incubation, hepatitis B e antigen （HBeAg） and hepatitis B surface antigen （HBsAg） of each group were assayed with ELISA and the survival rate of cells in each group was determined with 3- （ 4,5-dimethyhhiazol-2-yl ） -2,5-diphenyhe-trazolium bromide （ MTY ） reduction assay. Results ^125I-TFO showed a high stability with a radiolabeling rate of 〉93%. The radiochemical purity of labeled compound was 90.8%, 81.1% and 73.2% respectively after 12,48 and 72 h at 37℃. The peak inhibition effect of ^125I-TFO on synthesizing HBsAg and HBeAg by HepG2.2.15 cells were found at 48 h after transfection, with significantly the highest inhibition rate of 45.2% for HBsAg and 74.5% for HBeAg expression among the three groups （ P 〈 0.01 ）. As the transfection time prolonged its inhibition effects were stronger. Conclusion ^125I-TFO may inhibit the antigen expression of HBV and the growth of hepatocarcinoma cells, thus it may provide a new approach to develop gene-based radiotherapeutic pharmaceuticals for anti-HBV and HCC.

关 键 词：肝细胞瘤 肿瘤细胞 培养的 寡核苷酸类 碘放射性同位素 同位素标记 放射疗法 

分 类 号：R735.7[医药卫生—肿瘤]