Jansen B, Wadl H, Inoue S A, Trülzsch B, Selzer E, Duchêne M, Eichler H G, Wolff K, Pehamberger H
Institute for Specific Prophylaxis and Tropical Medicine, Vienna, Austria.
Antisense Res Dev. 1995 Winter;5(4):271-7. doi: 10.1089/ard.1995.5.271.
In our efforts to investigate the biologic role of Ha-ras oncogenes in human melanoma by Ha-ras phosphorothioate antisense oligonucleotides, we observed that antisense, sense, and scrambled control oligonucleotides at a concentration of 10 microM all similarly and strongly inhibited growth of our human melanoma target cell line SK-2 in vitro but without specific decrease of the target protein. Cell numbers with respect to the untreated control were reduced by 84% +/- 4.2% (ISD), 82.9% +/- 3.6%, and 84% +/- 3%, respectively. In vivo studies in a SCID-hu mouse model confirmed these findings. Both antisense and sense control oligonucleotides administered through osmotic pumps significantly (p < 0.006) reduced the mean tumor weight (1.5 g +/- 0.4 g and 1.8 g +/- 0.8 g, respectively) in comparison with saline-treated (5.7 g +/- 0.7 g) or untreated control animals (5.8 g +/- 1.0 g). The vascularity of oligonucleotide-treated tumors was greatly reduced. Clinical signs of oligonucleotide-related toxicity were not observed, and there was no evidence of histopathologic alterations in a variety of mouse tissues. We could demonstrate that the antimelanoma effects can be abrogated in vitro by adding basic fibroblast growth factor (bFGF). In the context of the importance of bFGF in melanocyte biology and angiogenesis, we argue in favor of an interaction between polyanionic phosphorothioate oligonucleotides and bFGF in our melanoma system. These findings stress the notion that phosphorothioate oligonucleotides may be promising antineoplastic lead compounds capable of employing antitumor effects by mechanisms other than specific inhibition of gene expression.
在我们通过Ha-ras硫代磷酸酯反义寡核苷酸研究Ha-ras癌基因在人类黑色素瘤中的生物学作用的过程中，我们观察到浓度为10微摩尔的反义、正义和随机对照寡核苷酸在体外均同样强烈地抑制了我们的人类黑色素瘤靶细胞系SK-2的生长，但靶蛋白并未特异性减少。相对于未处理的对照，细胞数量分别减少了84%±4.2%（平均标准差）、82.9%±3.6%和84%±3%。在SCID-hu小鼠模型中的体内研究证实了这些发现。与盐水处理组（5.7克±0.7克）或未处理的对照动物（5.8克±1.0克）相比，通过渗透泵给予的反义寡核苷酸和正义对照寡核苷酸均显著（p<0.006）降低了平均肿瘤重量（分别为1.5克±0.4克和1.8克±0.8克）。经寡核苷酸处理的肿瘤的血管生成大大减少。未观察到与寡核苷酸相关的毒性的临床体征，并且在多种小鼠组织中没有组织病理学改变的证据。我们能够证明，通过添加碱性成纤维细胞生长因子（bFGF），体外抗黑色素瘤作用可以被消除。鉴于bFGF在黑素细胞生物学和血管生成中的重要性，我们认为在我们的黑色素瘤系统中，聚阴离子硫代磷酸酯寡核苷酸与bFGF之间存在相互作用。这些发现强调了这样一种观点，即硫代磷酸酯寡核苷酸可能是有前景的抗肿瘤先导化合物，能够通过特异性抑制基因表达以外的机制发挥抗肿瘤作用。
