Department of Cell and Developmental Biology (formerly Anatomy and Developmental Biology), University College London, London, UK.
Eur J Cell Biol. 2011 Apr;90(4):312-22. doi: 10.1016/j.ejcb.2010.09.007. Epub 2010 Nov 18.
Intercellular communication via GAP Junctions plays an important role in tissue homeostasis, apoptosis, carcinogenesis, cell proliferation and differentiation. Hepatocyte connexins (Cx) 26 and 32 levels are decreased during the de-differentiation process of primary hepatocytes in culture, a situation that is also characterized by a decrease in S-Adenosylmethionine (SAMe) levels. In this current study, we show that SAMe supplementation in cultured hepatocytes every 12h, leads to an up-regulation of Cx26 and 32 mRNA and protein levels and blocks culture-induced Cx43 expression, although it failed to increase Cx26 and 32 membrane localization and GAP junction intracellular communication. SAMe reduced nuclear β-catenin accumulation, which is known to stimulate the TCF/LEF-dependent gene transcription of Cx43. Moreover SAMe-induced reduction in Cx43 and β-catenin was prevented by the proteasome inhibitor MG132, and was not mediated by GSK3 activity. SAMe, and its metabolite 5'-methylthioadenosine (MTA) increased Cx26 mRNA in a process partially mediated by Adenosine A(2A) receptors but independent of PKA. Finally livers from MAT1A knockout mice, characterized by low hepatic SAMe levels, express higher Cx43 and lower Cx26 and 32 protein levels than control mice. These results suggest that SAMe maintains a characteristic expression pattern of the different Cxs in hepatocytes by differentially regulating their levels.
细胞间隙连接通过缝隙连接进行细胞间通讯在组织稳态、细胞凋亡、癌变、细胞增殖和分化中发挥重要作用。原代培养肝细胞在去分化过程中,肝细胞连接蛋白(Cx)26 和 32 的水平降低,同时 S-腺苷甲硫氨酸(SAMe)水平也降低。在本研究中,我们发现每隔 12 小时向培养的肝细胞中补充 SAMe,可上调 Cx26 和 32 的 mRNA 和蛋白水平,并阻断培养诱导的 Cx43 表达,尽管它未能增加 Cx26 和 32 的膜定位和缝隙连接细胞内通讯。SAMe 减少了核内 β-连环蛋白的积累,已知β-连环蛋白会刺激 Cx43 的 TCF/LEF 依赖性基因转录。此外,SAMe 诱导的 Cx43 和 β-连环蛋白减少可被蛋白酶体抑制剂 MG132 阻止,且不通过 GSK3 活性介导。SAMe 及其代谢物 5'-甲基硫代腺苷(MTA)在部分通过腺苷 A(2A)受体介导、但不依赖 PKA 的过程中增加 Cx26 mRNA。最后,MAT1A 敲除小鼠肝脏中 SAMe 水平较低,其 Cx43 蛋白水平较高,Cx26 和 32 蛋白水平较低,与对照小鼠相比。这些结果表明,SAMe 通过差异调节不同 Cx 的水平,维持肝细胞中不同 Cx 的特征表达模式。