Nakayama M, Takahashi K, Hara E, Murakami O, Totsune K, Sone M, Satoh F, Shibahara S
Department of Molecular Biology and Applied Physiology, Tohoku University School of Medicine, Sendai, Japan.
J Cardiovasc Pharmacol. 1998;31 Suppl 1:S534-6. doi: 10.1097/00005344-199800001-00154.
Production and secretion of endothelin-1 (ET-1) and adrenomedullin (ADM) by a cultured human colorectal adenocarcinoma cell line, DLD-1, were studied by radioimmunoassay and Northern blot analysis. Both immunoreactive (IR)-ET and IR-ADM were detected by radioimmunoassay in the culture medium of DLD-1 (IR-ET 0.86 +/- 0.05 fmol/10(5) cells/ 24 h; IR-ADM 1.20 +/- 0.09 fmol/10(5) cells/24 h; n = 5, mean +/- SEM). An analysis by reverse-phase high-performance liquid chromatography (HPLC) of the IR-ET in the culture medium showed a major immunoreactive peak in the position of ET-1. Reverse-phase HPLC of the IR-ADM in the medium showed three immunoreactive peaks, one of which eluted in the position of human ADM. Northern blot analysis showed the expression of ET-1 mRNA and ADM mRNA in the DLD-1 cells. Treatment with interferon-gamma (1-100 U/ml) for 24 h decreased the IR-ET levels in the culture medium but significantly increased IR-ADM levels. This study has shown the production and secretion of two vasoactive peptides, ET-1 and ADM, by DLD-1 colorectal adenocarcinoma cells. The secretion of IR-ET was decreased by treatment with interferon-gamma. These findings suggest possible pathophysiologic roles for ET-1 and ADM in colon mucosal epithelial cells and tumors derived from them.
采用放射免疫分析法和Northern印迹分析法，研究了人结肠直肠腺癌细胞系DLD-1中内皮素-1（ET-1）和肾上腺髓质素（ADM）的产生与分泌。通过放射免疫分析法在DLD-1的培养基中检测到免疫反应性（IR）-ET和IR-ADM（IR-ET 0.86±0.05 fmol/10⁵细胞/24小时；IR-ADM 1.20±0.09 fmol/10⁵细胞/24小时；n = 5，平均值±标准误）。对培养基中的IR-ET进行反相高效液相色谱（HPLC）分析，结果显示在ET-1的位置有一个主要的免疫反应峰。对培养基中的IR-ADM进行反相HPLC分析，结果显示有三个免疫反应峰，其中一个在人ADM的位置洗脱。Northern印迹分析显示DLD-1细胞中ET-1 mRNA和ADM mRNA的表达。用干扰素-γ（1 - 100 U/ml）处理24小时可降低培养基中的IR-ET水平，但显著提高IR-ADM水平。本研究表明DLD-1结肠直肠腺癌细胞可产生和分泌两种血管活性肽ET-1和ADM。用干扰素-γ处理可降低IR-ET的分泌。这些发现提示ET-1和ADM在结肠黏膜上皮细胞及其衍生肿瘤中可能具有病理生理作用。
