Normanno N, Bianco C, Damiano V, de Angelis E, Selvam M P, Grassi M, Magliulo G, Tortora G, Bianco A R, Mendelsohn J, Salomon D S, Ciardiello F
Oncologia Sperimentale D, ITN-Fondazione Pascale and Cattedra di Clinica, Facoltá di Medicina e Chirurgia, Universitá Federico II, 80131 Naples, Italy.
Clin Cancer Res. 1996 Mar;2(3):601-9.
GEO is a well-differentiated colon cancer cell line that coexpresses the epidermal growth factor-like growth factors CRIPTO (CR), amphiregulin (AR), and transforming growth factor alpha (TGF-alpha). Antisense 20-mer phosphorothioate oligodeoxynucleotides (AS S-oligos) directed against CR, AR, and TGF-alpha mRNAs were equipotent in their ability to inhibit both the anchorage-dependent growth and the anchorage-independent growth (AIG) of GEO cells, with a 50% inhibitory concentration of about 5 micrometer in the AIG assay. A supraadditive effect was observed when a combination of S-oligos was used. For example, a combination of two different AS S-oligos (either AR + CR, or TGF-alpha + CR, or TGF-alpha + AR) at a concentration of 1 micrometer each (total concentration, 2 micrometer) resulted in 50% inhibition of GEO cells AIG, whereas the use of each AS S-Oligo at a 1 or 2 micrometer concentration resulted respectively in about 10 and 20% growth inhibition. A combination of the three AS S-oligos was even more effective, resulting in about 60% inhibition of GEO cells AIG at a concentration of 1 micrometer each (3 micrometer total concentration). The AS S-oligos were also able to inhibit specifically the expression of either AR, CR, or TGF-alpha proteins in GEO cells, as assessed using immunocytochemistry or Western blot analysis. Finally, a supraadditive growth inhibitory effect of the AS S-oligos and an epidermal growth factor receptor-blocking antibody (monoclonal antibody 528) was observed. These data suggest that the use of a combination of AS S-oligos directed against different growth factors and antibodies directed against their receptors might result in an efficient inhibition of colon carcinoma cell growth.
GEO是一种高分化结肠癌细胞系，共表达表皮生长因子样生长因子CRIPTO（CR）、双调蛋白（AR）和转化生长因子α（TGF-α）。针对CR、AR和TGF-α mRNA的反义20聚硫代磷酸酯寡脱氧核苷酸（AS S-寡核苷酸）在抑制GEO细胞的贴壁依赖性生长和贴壁非依赖性生长（AIG）方面具有同等效力，在AIG试验中50%抑制浓度约为5微摩尔。当使用S-寡核苷酸组合时观察到超加成效应。例如，两种不同的AS S-寡核苷酸（AR + CR，或TGF-α + CR，或TGF-α + AR）各以1微摩尔的浓度（总浓度为2微摩尔）组合，可导致GEO细胞AIG受到50%的抑制，而每种AS S-寡核苷酸以1或2微摩尔浓度使用时分别导致约10%和20%的生长抑制。三种AS S-寡核苷酸的组合甚至更有效，各以1微摩尔的浓度（总浓度为3微摩尔）可导致GEO细胞AIG受到约60%的抑制。如使用免疫细胞化学或蛋白质印迹分析所评估的，AS S-寡核苷酸还能够特异性抑制GEO细胞中AR、CR或TGF-α蛋白的表达。最后，观察到AS S-寡核苷酸与表皮生长因子受体阻断抗体（单克隆抗体528）具有超加成生长抑制作用。这些数据表明，使用针对不同生长因子的AS S-寡核苷酸组合及其受体的抗体可能会有效抑制结肠癌细胞的生长。
