Massagué J
J Biol Chem. 1984 Aug 10;259(15):9756-61.
We have isolated a strongly mitogenic, type beta transforming growth factor (beta TGF) released by Snyder-Theilen feline sarcoma virus-transformed rat embryo (FeSV-Fre) cells that induces phenotypic transformation of normal NRK cells when they are concomitantly stimulated by analogues of epidermal growth factor (EGF). Molecule filtration chromatography separates beta TGF from an EGF-like TGF (eTGF) which is also present in acid extracts from medium conditioned by FeSV-Fre cells (J. Massagué, (1983) J. Biol. Chem. 258, 13606-13613). Final purification of beta TGF is achieved by reverse phase high pressure liquid chromatography (HPLC) on octadecyl support, molecular filtration HPLC, and nonreducing dodecyl sulfate-polyacrylamide gel electrophoresis steps, yielding a 300,000-fold purified polypeptide with a final recovery of 21%. The purified rat beta TGF consists of two Mr = 11,000-12,000 polypeptide chains disulfide-linked as a Mr = 23,000 dimer. Induction of anchorage-independent proliferation of NRK cells by rat beta TGF depends on the simultaneous presence of eTGF or EGF. In the presence of a saturating (300 pM) concentration of either rat eTGF or mouse EGF, half-maximal anchorage-independent proliferation of NRK cells is obtained with 4-6 pM rat beta TGF. In the presence of a saturating (20 pM) concentration of rat beta TGF, half-maximal anchorage-independent proliferation of NRK cells is obtained with either rat eTGF or mouse EGF at a 50-70 pM concentration. Rat beta TGF is also able to induce DNA synthesis and cell proliferation on growth-arrested NRK, human lung, and Swiss mouse 3T3 fibroblast monolayers, this effect being half-maximal at 2-3 pM beta TGF for NRK cells. These results identify eTGF and beta TGF as the two synergistically acting factors responsible for the transforming action of culture fluids from FeSV-Fre cells.
我们从斯奈德 - 泰伦猫肉瘤病毒转化的大鼠胚胎（FeSV - Fre）细胞中分离出一种具有强促有丝分裂作用的β型转化生长因子（β - TGF），当正常NRK细胞同时受到表皮生长因子（EGF）类似物刺激时，该因子可诱导其发生表型转化。分子过滤色谱法可将β - TGF与一种也存在于FeSV - Fre细胞条件培养基酸提取物中的类表皮生长因子（eTGF）分离（J. 马萨格，（1983年）《生物化学杂志》258卷，13606 - 13613页）。β - TGF的最终纯化通过在十八烷基支持物上进行反相高压液相色谱（HPLC）、分子过滤HPLC以及非还原十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳步骤实现，得到一种纯化了300,000倍的多肽，最终回收率为21%。纯化的大鼠β - TGF由两条Mr = 11,000 - 12,000的多肽链通过二硫键连接形成Mr = 23,000的二聚体。大鼠β - TGF诱导NRK细胞非贴壁依赖性增殖取决于eTGF或EGF的同时存在。在存在饱和浓度（300 pM）的大鼠eTGF或小鼠EGF的情况下，4 - 6 pM的大鼠β - TGF可使NRK细胞实现半最大非贴壁依赖性增殖。在存在饱和浓度（20 pM）的大鼠β - TGF的情况下，50 - 70 pM浓度的大鼠eTGF或小鼠EGF可使NRK细胞实现半最大非贴壁依赖性增殖。大鼠β - TGF还能够诱导生长停滞的NRK、人肺和瑞士小鼠3T3成纤维细胞单层进行DNA合成和细胞增殖，对于NRK细胞而言，这种效应在2 - 3 pM的β - TGF时达到半最大。这些结果表明，eTGF和β - TGF是两种协同作用的因子，它们共同导致了FeSV - Fre细胞培养液的转化作用。
