Murakami A, Yamayoshi A, Iwase R, Nishida J, Yamaoka T, Wake N
Department of Polymer Science and Engineering, Kyoto Institute of Technology, Matsugasaki, 606-8585, Kyoto, Japan.
Eur J Pharm Sci. 2001 Apr;13(1):25-34. doi: 10.1016/s0928-0987(00)00204-9.
The antisense strategy has been applied to regulate gene expression in a sequence specific manner, which enables suppression of the proliferation of cancer cells and exploration of the functions of unknown genes. In order to generalize and to enhance the ability of the strategy, functionalization of antisense DNAs was done using a photo-crosslinking reagent, 4,5',8-trimethylpsoralen, and the possibility of photodynamic antisense regulation of gene expression was examined. Psoralen-conjugated oligo(nucleoside phosphorothioate)s (Ps-S-oligo) were prepared and used to inhibit the proliferation of human cervical carcinoma cells. Upon UVA irradiation of Ps-S-oligo treated cells, Ps-S-oligo complementary to the initiation codon region (Ps-P-As) of HPV18-E6*-mRNA of human cervical carcinoma cells inhibited drastically the cell growth (IC(50)=16 nM). In contrast, Ps-S-oligo with mismatched sequences and scrambled one showed lesser inhibitory effects than Ps-P-As. These results showed that the inhibition by Ps-S-oligo was dependent on (a) sequence, (b) UVA irradiation, (c) concentration and (d) cell line. The amount of intact HPV18-E6*-mRNA was decreased in a sequence dependent manner, indicating that the antiproliferative effect of Ps-P-As was an antisense manner. The psoralen-conjugated antisense DNA has significant potential to regulate gene expression, which may provide useful information to explore the novel gene regulating reagents.
反义策略已被应用于以序列特异性方式调节基因表达，这使得能够抑制癌细胞增殖并探索未知基因的功能。为了推广并增强该策略的能力，使用光交联试剂4,5',8-三甲基补骨脂素对反义DNA进行功能化，并研究了基因表达的光动力反义调节的可能性。制备了补骨脂素缀合的寡聚（核苷硫代磷酸酯）（Ps-S-寡聚体）并用于抑制人宫颈癌细胞的增殖。在用Ps-S-寡聚体处理的细胞进行UVA照射后，与人宫颈癌细胞HPV18-E6 * -mRNA的起始密码子区域（Ps-P-As）互补的Ps-S-寡聚体极大地抑制了细胞生长（IC(50)=16 nM）。相比之下，具有错配序列和随机序列的Ps-S-寡聚体显示出比Ps-P-As更小的抑制作用。这些结果表明，Ps-S-寡聚体的抑制作用取决于（a）序列、（b）UVA照射、（c）浓度和（d）细胞系。完整的HPV18-E6 * -mRNA的量以序列依赖性方式减少，表明Ps-P-As的抗增殖作用是反义方式。补骨脂素缀合的反义DNA具有调节基因表达的巨大潜力，这可能为探索新型基因调节试剂提供有用信息。
