Suzuki N, Matsumoto H, Kitada C, Kimura S, Fujino M
Tsukuba Research Laboratories, Takeda Chemical Industries, Ltd., Ibaraki.
J Biochem. 1989 Nov;106(5):736-41. doi: 10.1093/oxfordjournals.jbchem.a122925.
Immunoreactive endothelin (ET) and big-endothelin (big-ET) in conditioned media of endothelial and of non-endothelial cells were studied using sandwich-type enzyme immunoassays. Immunoreactivities of both ETs were detected in the media of all four endothelial cells tested. Among non-endothelial cells, tumor cell lines with epithelial-like morphology also produced immunoreactive ET and/or big-ET, although the total amount of ETs was one or two orders of magnitude less than that produced by PAE (porcine aortic endothelial cells). Immunoreactive ETs produced by HepG-2 (human hepatocellular carcinoma) and some other cells were characterized by gel-filtration HPLC and reverse-phase HPLC. These studies revealed the production of ET-1 and human big-ET-1 by these cells, although the immunoreactive ETs produced by the tumor cells were more heterogeneous than those produced by endothelial cells. The regulatory effects of thrombin and transforming growth factor-beta (TGF-beta) on the production of ETs were investigated. TGF-beta markedly stimulated the production of both ETs in HepG-2 and slightly decreased the big-ET level in A549 (human lung carcinoma). HPLC analysis showed that the major immunoreactive ETs induced by TGF-beta in HepG-2 were identical to ET-1 and human big-ET-1. These results demonstrated that production of ET-1 and big-ET-1 was not restricted to endothelial cells and was induced by TGF-beta in HepG-2 at the same levels as those produced by PAE.
