^(125)碘标记三链形成寡核苷酸对肝癌细胞中病毒复制和细胞生长的抑制作用  

Inhibition effects of ^(125)I-antigene oligonucleotide against hepatitis B virus duplication and cell growth in hepatoma carcinoma cells

作　　者：李丹[1] 侯敏[1] 吕中伟[1] 吕明丽[1] 蔡海东[1] 余飞[1] 孙明[1] 王舰[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

出　　处：《上海医学》2010年第3期290-294,共5页Shanghai Medical Journal

基　　金：国家自然科学基金项目资助(30901770)

摘　　要：目的探讨125碘(I)标记三链形成寡核苷酸(TFO)与乙型肝炎病毒(HBV)DNA的特异性结合能力及其对肝癌细胞HepG2.2.15中病毒复制及细胞生长的抑制作用。方法合成能与HBV前S基因特异结合的TFO,并以125I标记。将HepG2.2.15细胞分别与125I-TFO实验组(125I-TFO组)、等量TFO对照组(TFO组)、相同放射性活度的钠125碘(Na125I)组和空白对照组共同培养,荧光定量逆转录-聚合酶链反应(RT-PCR)检测细胞培养液中HBVDNA拷贝量变化,采用四甲基偶氮唑蓝(MTT)比色法检测转染后各组细胞的存活率。结果RT-PCR显示转染48h后,4组间病毒拷贝数量的差异有统计学意义(P<0.01),125I-TFO组对病毒复制的抑制作用显著强于空白对照组(P<0.01)、125I对照组(P<0.01)和TFO组(P<0.01);TFO组的抑制病毒复制作用显著强于空白对照组(P<0.05)。转染24、48、72h,125I-TFO组对HepG2.2.15细胞的生长抑制作用显著高于其他3组(P值均<0.01),并随时间的增加125I-TFO对细胞生长的抑制作用呈线性上升趋势。结论125I-TFO可有效地抑制肝癌细胞中HBV的复制及肝癌细胞的生长,为今后制备抗HBV、抗HBV相关肝细胞癌的放射性基因治疗药物奠定了基础。Objective To examine the binding ability of 125I labeled-triplex forming oligonucleotide （125I -TFO） to the pre S gene of hepatitis B virus（HBV）DNA and examine the inhibitory effect of 125I-TFO on HBV duplication and cell growth of hepatoma carcinoma cells. Methods TFO complementary to pre S gene of HBV was synthesized and labeled with 125I. Then Hep G2.2.15 cells were divided into four groups： 125I-TFO group, TFO group, Na 125I group and blank control group. The culture supernatants in the four groups were collected after 24, 48, and 72 h. The virus DNA copies were examined by real-time PCR（RT-PCR）, and the survival rate of cells in each group was assayed by MTT reduction assay. Results The HBV DNA copies of the four groups were significantly different 48 h after transfection （P0.01）. The duplication of HBV DNA in 125I-TFO group was significantly inhibited compared with the blank control group （P0.01）, 125I group （P0.01） and TFO group （P0.01）; and that of the TFO group was also significantly different from that of the blank control group （P0.05）. 125I-TFO group showed a significantly stronger inhibitory effect on HepG2.2.15 cell growth compared with other groups at 24 , 48 and 72 h after transfection （P0.01）, and the inhibitory effect increased linearly with the time. Conclusion It is concluded that 125I-TFO may inhibit the antigen expression of HBV in hepatic celluar carcinoma （HCC） cells and induce cell death of HCC cells, which paves a way for developing gene-based radiotherapy pharmaceuticals for anti-HBV and HCC.

关 键 词：三链形成寡核苷酸 HEPG2.2.15细胞 125碘 

分 类 号：R735.7[医药卫生—肿瘤]