甲胎蛋白反义寡脱氧核苷酸与5-氟尿嘧啶对人肝癌细胞生长的联合作用

Combined effect of alpha-fetoprotein antisense oligodeoxy-nucleotides and 5-fluorouracil on human hepatoma cell growth.

作者信息

Wang X, Zhang R, Xie H

机构信息

Department of Biotherapy, Shanghai Institute of Cell Biology, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.

出版信息

Chin Med J (Engl). 1999 Aug;112(8):743-6.

PMID:11601286

Abstract

OBJECTIVE

To investigate the effect of alpha-fetoprotein (AFP) antisense phosphorothioate oligodeoxynucleotides in combination with 5-fluorouracil (5-FU) on the growth of BEL-7404 human hepatoma cells in vitro.

METHODS

Phosphorothioate oligodeoxynucleotides were synthesized by beta-cyanoethyl-phosphoramidite chemistry. AFP gene expression in human hepatoma cells was determined by avidin-biotin-peroxidase complex (ABC) immunocytochemical method. Cell growth in the presence or absence of experimental agents was measured using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-dipheny ltetrazolium (MTT) microculture tetrazolium assay.

RESULTS

AFP antisense oligomers markedly suppressed the growth of BEL-7404 human hepatoma cells in vitro by sequence-specific blocking of the AFP gene expression in the cells (P < 0.05). 5-FU also inhibited the hepatoma cell growth in a dose-dependent manner when used alone (P < 0.05). The combined treatment with AFP antisense oligomers and 5-FU showed significantly enhanced hepatoma cell growth inhibition than either AFP antisense or 5-FU treated cells alone (P < 0.05).

CONCLUSION

Combined use of AFP antisense oligomers and 5-FU could more effectively inhibit the growth of BEL-7404 human hepatoma cells in vitro.

摘要

目的

研究α-甲胎蛋白（AFP）反义硫代磷酸酯寡脱氧核苷酸与5-氟尿嘧啶（5-FU）联合应用对人肝癌BEL-7404细胞体外生长的影响。

方法

采用β-氰乙基亚磷酰胺化学合成硫代磷酸酯寡脱氧核苷酸。用抗生物素蛋白-生物素-过氧化物酶复合物（ABC）免疫细胞化学方法检测人肝癌细胞中AFP基因的表达。采用3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐（MTT）微量培养四氮唑法检测在有无实验药物存在时细胞的生长情况。

结果

AFP反义寡聚体通过序列特异性阻断细胞中AFP基因的表达，显著抑制了人肝癌BEL-7404细胞的体外生长（P<0.05）。单独使用5-FU时也能以剂量依赖方式抑制肝癌细胞生长（P<0.05）。与单独使用AFP反义寡聚体或5-FU处理的细胞相比，AFP反义寡聚体与5-FU联合处理对肝癌细胞生长的抑制作用显著增强（P<0.05）。