Zhang G H, Ichimura T, Wallin A, Kan M, Stevens J L
W. Alton Jones Cell Science Center, Lake Placid, New York 12946.
J Cell Physiol. 1991 Aug;148(2):295-305. doi: 10.1002/jcp.1041480216.
Growth factors may play an important role in regulating the growth of the proximal tubule epithelium. To determine which growth factors could be involved, we have investigated the mitogenicity of various purified factors in rat kidney proximal tubule epithelial (RPTE) cells cultured in defined medium. Fibroblast growth factors, aFGF (acidic FGF) and bFGF (basic FGF), stimulate DNA synthesis in a dose-dependent manner, with ED50 values of 4.5 and 3.2 ng/ml, respectively; their effects are not additive. With cholera toxin in the medium, both aFGF and bFGF can replace insulin or epidermal growth factor (EGF) to attain the maximum level of cell growth, but they cannot replace cholera toxin. Cholera toxin specifically potentiates the effects of FGFs on DNA synthesis. At high cell density, both insulin and insulin-like growth factor 1 (IGF-1) induce DNA synthesis more effectively than EGF, FGFs and cholera toxin. The high concentration (0.2-1.0 microgram/ml) of insulin required for cell growth can be replaced by a low concentration of IGF-1 (10-20 ng/ml), indicating that insulin probably acts through a low affinity interaction with the IGF-1 receptor. Transforming growth factor-beta 1 (TGF-beta 1) inhibits DNA synthesis induced by individual factors and combinations of factors in a concentration-dependent manner. Northern blot analysis shows that mRNA for TGF-beta 1, IGF-1, and aFGF, but not bFGF are present in rat kidney. Western blot analysis and bioassay data confirmed that the majority of FGF-like protein in rat kidney is aFGF. The data suggest that in addition to EGF, IGFs, and TGF-beta, FGFs may also be important kidney-derived regulators of proximal tubule epithelial cell growth in vivo and in vitro.
生长因子可能在调节近端肾小管上皮细胞的生长中发挥重要作用。为了确定哪些生长因子可能参与其中,我们研究了在限定培养基中培养的大鼠肾近端小管上皮(RPTE)细胞中各种纯化因子的促有丝分裂活性。成纤维细胞生长因子,即酸性成纤维细胞生长因子(aFGF)和碱性成纤维细胞生长因子(bFGF),以剂量依赖方式刺激DNA合成,ED50值分别为4.5和3.2 ng/ml;它们的作用并非相加性的。培养基中加入霍乱毒素后,aFGF和bFGF均可替代胰岛素或表皮生长因子(EGF)达到细胞生长的最大水平,但它们不能替代霍乱毒素。霍乱毒素能特异性增强FGFs对DNA合成的作用。在高细胞密度下,胰岛素和胰岛素样生长因子1(IGF-1)诱导DNA合成的效果比EGF、FGFs和霍乱毒素更有效。细胞生长所需的高浓度胰岛素(0.2 - 1.0微克/毫升)可被低浓度的IGF-1(10 - 20 ng/ml)替代,这表明胰岛素可能通过与IGF-1受体的低亲和力相互作用发挥作用。转化生长因子-β1(TGF-β1)以浓度依赖方式抑制单个因子及因子组合诱导的DNA合成。Northern印迹分析表明,大鼠肾脏中存在TGF-β1、IGF-1和aFGF的mRNA,但不存在bFGF的mRNA。蛋白质印迹分析和生物测定数据证实,大鼠肾脏中大多数FGF样蛋白是aFGF。这些数据表明,除了EGF、IGFs和TGF-β外,FGFs在体内和体外也可能是肾脏来源的近端小管上皮细胞生长的重要调节因子。