Johnson B J, White M E, Hathaway M R, Dayton W R
Department of Animal Science, University of Minnesota, St. Paul, USA.
J Cell Physiol. 1999 May;179(2):237-43. doi: 10.1002/(SICI)1097-4652(199905)179:2<237::AID-JCP15>3.0.CO;2-G.
Insulin-like growth factor binding proteins (IGFBPs) affect the biological activity of IGF-I in several cell types, including cultured muscle cells. Additionally, at least one of the IGFBPs, IGFBP-3, has been shown to have IGF-independent effects on cell proliferation. Numerous studies have shown that immortalized muscle cell lines produce various IGFBPs, but to date no muscle cell line has been reported to produce IGFBP-3 protein or mRNA. Unlike muscle cell lines, primary cultures of porcine embryonic myogenic cells express IGFBP-3 mRNA and secrete a protein that is immunologically identifiable as IGFBP-3. Additionally, steady-state IGFBP-3 levels change significantly during differentiation. Here we report that differentiation of porcine myogenic cells in an IGFBP-3-free medium is accompanied by reduced steady-state IGFBP-3 mRNA levels. Steady-state levels of IGFBP-3 mRNA decreased approximately sevenfold (P < .05) during differentiation and then increased to predifferentiation levels once differentiation was complete. Addition of TGF-beta1 (0.5 ng/ml) to porcine myogenic cultures suppressed fusion and resulted in a sevenfold increase in steady-state IGFBP-3 mRNA and a 1.8-fold increase in IGFBP-3 protein levels as compared to untreated control cultures (P < .05). Results suggest that alterations in IGFBP-3 mRNA and protein may play a role in differentiation of porcine embryonic muscle cells.
胰岛素样生长因子结合蛋白(IGFBPs)在多种细胞类型中影响IGF-I的生物学活性,包括培养的肌肉细胞。此外,至少有一种IGFBPs,即IGFBP-3,已被证明对细胞增殖具有不依赖IGF的作用。大量研究表明,永生化肌肉细胞系会产生多种IGFBPs,但迄今为止,尚无报道称有肌肉细胞系能产生IGFBP-3蛋白或mRNA。与肌肉细胞系不同,猪胚胎成肌细胞的原代培养物表达IGFBP-3 mRNA,并分泌一种在免疫学上可鉴定为IGFBP-3的蛋白质。此外,在分化过程中,IGFBP-3的稳态水平会发生显著变化。在此我们报告,在无IGFBP-3的培养基中猪成肌细胞的分化伴随着IGFBP-3 mRNA稳态水平的降低。在分化过程中,IGFBP-3 mRNA的稳态水平下降了约7倍(P <.05),一旦分化完成,又会升至分化前水平。与未处理的对照培养物相比,向猪成肌细胞培养物中添加TGF-β1(0.5 ng/ml)会抑制融合,并导致IGFBP-3 mRNA稳态水平增加7倍,IGFBP-3蛋白水平增加1.8倍(P <.05)。结果表明,IGFBP-3 mRNA和蛋白的改变可能在猪胚胎肌肉细胞的分化中起作用。