Lobb R R, Rybak S M, St Clair D K, Fett J W
Biochem Biophys Res Commun. 1986 Sep 30;139(3):861-7. doi: 10.1016/s0006-291x(86)80257-1.
Cell lysates of two established human tumor lines, a medulloblastoma (TE671), and a rhabdomyosarcoma (RD), contain mitogenic activity which elutes from heparin-Sepharose under conditions typical of class 1 heparin-binding growth factors, such as acidic brain fibroblast growth factor. The presence of this class of mitogen in both cell lines was confirmed by their chromatographic behavior on reversed-phase C3 columns, and by the ability of heparin to enhance their mitogenic activity. Using a specific synthetic DNA probe, RNA's were isolated from both cell lines by hybridization-selection, translated in vitro, and translated proteins affinity fractionated on heparin-Sepharose. The results demonstrate that TE671 and RD cell lysates contain mRNA's for mitogens related to acidic brain fibroblast growth factor, and also suggest that high molecular weight proteins exist that are closely related to, or are precursor forms of, the class 1 mitogens.
两种已建立的人类肿瘤细胞系,即髓母细胞瘤(TE671)和横纹肌肉瘤(RD)的细胞裂解物含有促有丝分裂活性,这种活性在典型的1类肝素结合生长因子(如酸性脑成纤维细胞生长因子)的条件下从肝素-琼脂糖凝胶中洗脱出来。这两类细胞系中这类有丝分裂原的存在通过它们在反相C3柱上的色谱行为以及肝素增强其促有丝分裂活性的能力得到证实。使用一种特异性合成DNA探针,通过杂交选择从这两类细胞系中分离出RNA,在体外进行翻译,并将翻译后的蛋白质在肝素-琼脂糖凝胶上进行亲和分级分离。结果表明,TE671和RD细胞裂解物含有与酸性脑成纤维细胞生长因子相关的有丝分裂原的mRNA,并且还表明存在与1类有丝分裂原密切相关或为其前体形式的高分子量蛋白质。