Hagiwara K, Kobayashi T, Tobita M, Kikyo N, Yazaki Y, Okabe T
Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo.
Jpn J Cancer Res. 1995 Feb;86(2):202-7. doi: 10.1111/j.1349-7006.1995.tb03040.x.
We have found growth-promoting activity for vascular endothelial cells in the conditioned medium of a human lung cancer cell line, T3M-11. Purification and characterization of the growth-promoting activity have been carried out using ammonium sulfate precipitation and gel-exclusion chromatography. The activity migrated as a single peak just after ribonuclease. It did not bind to a heparin affinity column. These results suggest that the activity is not a heparin-binding growth factor (including fibroblast growth factors) or a vascular endothelial growth factor. To identify the molecule exhibiting the growth-promoting activity, a cDNA encoding the growth factor was isolated through functional expression cloning in COS-1 cells from a cDNA library prepared from T3M-11 cells. The nucleotide sequence encoded by the cDNA proved to be identical with that of insulin-like growth factor II.
我们在人肺癌细胞系T3M - 11的条件培养基中发现了对血管内皮细胞的促生长活性。利用硫酸铵沉淀和凝胶排阻色谱法对促生长活性进行了纯化和表征。该活性在核糖核酸酶之后以单一峰的形式迁移。它不与肝素亲和柱结合。这些结果表明，该活性不是肝素结合生长因子（包括成纤维细胞生长因子）或血管内皮生长因子。为了鉴定具有促生长活性的分子，通过功能表达克隆从T3M - 11细胞制备的cDNA文库中分离出了编码该生长因子的cDNA，该cDNA在COS - 1细胞中进行表达。结果证明，该cDNA编码的核苷酸序列与胰岛素样生长因子II的序列相同。
