Netzer P, Domek M, Pai R, Halter F, Tarnawski A
VA Medical Center, Long Beach, California, USA.
Aliment Pharmacol Ther. 2001 Oct;15(10):1673-9. doi: 10.1046/j.1365-2036.2001.01084.x.
Basic fibroblast growth factor has been shown to be mitogenic in colon cancer cell lines. In human malignant melanoma cells, antisense oligodeoxynucleotides targeted against basic fibroblast growth factor messenger RNA significantly inhibit cell growth. However, the efficacy of such an antisense oligodeoxynucleotide strategy has not been evaluated for colon cancer cells.
To investigate whether basic fibroblast growth factor can stimulate the growth of HT-29 human colon cancer cells and whether antisense oligodeoxynucleotides can inhibit growth of these cells at baseline.
Western blotting analyses were used to confirm the presence of basic fibroblast growth factor protein in this cell line. Cell growth was assessed after 2, 4 and 6 days of treatment by cell counting using the trypan blue exclusion method. Phosphorothioate-modified oligodeoxynucleotides (10 microM) were used, complementary to codon 60 of the basic fibroblast growth factor messenger RNA. Cationic liposomes (DOTAP) were used to enhance the cellular uptake of the oligodeoxynucleotides.
Western blotting demonstrated the presence of basic fibroblast growth factor protein in this cell line. Basic fibroblast growth factor (1-40 ng/mL) dose-dependently stimulated cell growth and peak values were obtained at a dose of 20 ng/mL. By contrast, antisense oligodeoxynucleotide treatment significantly inhibited cell growth compared with the sense oligodeoxynucleotide-treated cells (P=0.007). This inhibition was reversed by the addition of basic fibroblast growth factor, 20 ng/mL.
Treatment targeted against basic fibroblast growth factor messenger RNA inhibits growth of HT-29 human colon cancer cells. This finding may provide a rationale for the therapeutic use of antisense oligodeoxynucleotides targeted at basic fibroblast growth factor for the treatment of colon cancer.
碱性成纤维细胞生长因子已被证明在结肠癌细胞系中具有促有丝分裂作用。在人类恶性黑色素瘤细胞中，针对碱性成纤维细胞生长因子信使核糖核酸的反义寡脱氧核苷酸可显著抑制细胞生长。然而，尚未评估这种反义寡脱氧核苷酸策略对结肠癌细胞的疗效。
研究碱性成纤维细胞生长因子是否能刺激HT-29人结肠癌细胞的生长，以及反义寡脱氧核苷酸在基线时是否能抑制这些细胞的生长。
采用蛋白质免疫印迹分析来确认该细胞系中碱性成纤维细胞生长因子蛋白的存在。使用台盼蓝排斥法通过细胞计数在处理2、4和6天后评估细胞生长。使用与碱性成纤维细胞生长因子信使核糖核酸密码子60互补的硫代磷酸酯修饰的寡脱氧核苷酸（10微摩尔）。使用阳离子脂质体（DOTAP）来增强寡脱氧核苷酸的细胞摄取。
蛋白质免疫印迹证明该细胞系中存在碱性成纤维细胞生长因子蛋白。碱性成纤维细胞生长因子（1 - 40纳克/毫升）剂量依赖性地刺激细胞生长，在20纳克/毫升的剂量下获得峰值。相比之下，与正义寡脱氧核苷酸处理的细胞相比，反义寡脱氧核苷酸处理显著抑制细胞生长（P = 0.007）。加入20纳克/毫升的碱性成纤维细胞生长因子可逆转这种抑制作用。
针对碱性成纤维细胞生长因子信使核糖核酸的治疗可抑制HT-29人结肠癌细胞的生长。这一发现可能为使用针对碱性成纤维细胞生长因子的反义寡脱氧核苷酸治疗结肠癌提供理论依据。
