Riuzzi Francesca, Sorci Guglielmo, Donato Rosario
Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Casella Postale 81 Succursale 3, 06122 Perugia, Italy.
J Biol Chem. 2006 Mar 24;281(12):8242-53. doi: 10.1074/jbc.M509436200. Epub 2006 Jan 9.
We reported that RAGE (receptor for advanced glycation end products), a multiligand receptor of the immunoglobulin superfamily expressed in myoblasts, when activated by its ligand amphoterin (HMGB1), stimulates rat L6 myoblast differentiation via a Cdc42-Rac-MKK6-p38 mitogen-activated protein kinase pathway, and that RAGE expression in skeletal muscle tissue is developmentally regulated. We show here that inhibition of RAGE function via overexpression of a signaling deficient RAGE mutant (RAGE delta cyto) results in increased myoblast proliferation, migration, and invasiveness, and decreased apoptosis and adhesiveness, whereas myoblasts overexpressing RAGE behave the opposite, compared with mock-transfected myoblasts. These effects are accompanied by a decreased induction of the proliferation inhibitor, p21(Waf1), and increased induction of cyclin D1 and extent of Rb, ERK1/2, and JNK phosphorylation in L6/RAGE delta cyto myoblasts, the opposite occurring in L6/RAGE myoblasts. Neutralization of culture medium amphoterin negates effects of RAGE activation, suggesting that amphoterin is the RAGE ligand involved in RAGE-dependent effects in myoblasts. Finally, mice injected with L6/RAGE delta cyto myoblasts develop tumors as opposed to mice injected with L6/RAGE or L6/mock myoblasts that do not. Thus, the amphoterin/RAGE pair stimulates myoblast differentiation by the combined effect of stimulation of differentiation and inhibition of proliferation, and deregulation of RAGE expression in myoblasts might contribute to their neoplastic transformation.
我们曾报道,晚期糖基化终末产物受体(RAGE)是免疫球蛋白超家族的一种多配体受体,在成肌细胞中表达,当其被配体两性调节蛋白(HMGB1)激活时,可通过Cdc42-Rac-MKK6-p38丝裂原活化蛋白激酶途径刺激大鼠L6成肌细胞分化,且骨骼肌组织中的RAGE表达受发育调控。我们在此表明,通过过表达信号缺陷型RAGE突变体(RAGEδcyto)抑制RAGE功能会导致成肌细胞增殖、迁移和侵袭增加,凋亡和黏附减少,而与mock转染的成肌细胞相比,过表达RAGE的成肌细胞表现则相反。这些效应伴随着增殖抑制剂p21(Waf1)诱导减少,细胞周期蛋白D1诱导增加,以及L6/RAGEδcyto成肌细胞中Rb、ERK1/2和JNK磷酸化程度增加,而在L6/RAGE成肌细胞中情况则相反。中和培养基中的两性调节蛋白可消除RAGE激活的效应,这表明两性调节蛋白是参与成肌细胞中RAGE依赖性效应的RAGE配体。最后,注射L6/RAGEδcyto成肌细胞的小鼠会发生肿瘤,而注射L6/RAGE或L6/mock成肌细胞的小鼠则不会。因此,两性调节蛋白/RAGE对通过刺激分化和抑制增殖的联合作用刺激成肌细胞分化,而成肌细胞中RAGE表达失调可能有助于其肿瘤转化。