Ohkubo S, Ogi K, Hosoya M, Matsumoto H, Suzuki N, Kimura C, Ondo H, Fujino M
Tsukuba Research Laboratories, Takeda Chemical Industries, Ibaraki, Japan.
FEBS Lett. 1990 Nov 12;274(1-2):136-40. doi: 10.1016/0014-5793(90)81348-r.
Analysis of culture medium of human renal adenocarcinoma cells ACHN by RP-HPLC suggested that the cells specifically secreted human endothelin-2 (ET-2). cDNAs encoding human ET-2 precursor were cloned from a cDNA library constructed with mRNA derived from the ACHN cells, and the nucleotide and deduced amino acid sequences were determined. The ET-2 cDNA was revealed to contain 1.3 kb and encode prepro-ET-2 protein consisting of 178 amino acid residues. The ET-like sequences found in the prepro-ET-1 and -ET-3 was conserved in this prepro-ET-2. The Northern blot analysis of mRNA suggested that the transcript of ET-2 gene was 1.4 kb. This is the first direct evidence that human ET-2 gene was expressed specifically in tumor cells.
采用反相高效液相色谱法（RP-HPLC）对人肾腺癌细胞ACHN的培养基进行分析，结果表明这些细胞特异性分泌人内皮素-2（ET-2）。从用ACHN细胞来源的mRNA构建的cDNA文库中克隆出编码人ET-2前体的cDNA，并测定了其核苷酸和推导的氨基酸序列。结果显示ET-2 cDNA含有1.3 kb，编码由178个氨基酸残基组成的前内皮素原-2蛋白。在前内皮素原-1和-3中发现的ET样序列在该前内皮素原-2中保守。对mRNA的Northern印迹分析表明ET-2基因的转录本为1.4 kb。这是人类ET-2基因在肿瘤细胞中特异性表达的首个直接证据。
