Peura Matti, Bizik Jozef, Salmenperä Pertteli, Noro Ariel, Korhonen Matti, Pätilä Tommi, Vento Antti, Vaheri Antti, Alitalo Riitta, Vuola Jyrki, Harjula Ari, Kankuri Esko
Institute of Biomedicine, Pharmacology, Biomedicum, University of Helsinki, Helsinki, Finland.
Wound Repair Regen. 2009 Jul-Aug;17(4):569-77. doi: 10.1111/j.1524-475X.2009.00507.x.
We previously showed cell-cell contacts of human dermal fibroblasts to induce expression of the hepatocyte growth factor/scatter factor (HGF) in a process designated as nemosis. Now we report on nemosis initiation in bone marrow mesenchymal stem cells (BMSCs). Because BMSCs are being used increasingly in cell transplantation therapy we aimed to demonstrate a functional effect and benefit of BMSC nemosis for wound healing. Nemotic and monolayer cells were used to stimulate HaCaT keratinocyte migration in a scratch-wound healing assay. Both indicators of nemosis, HGF production and cyclooxygenase-2 expression, were induced in BMSC spheroids. When compared with a similar amount of cells as monolayer, nemotic cells induced keratinocyte in vitro scratch-wound healing in a concentration-dependent manner. The HGF receptor, c-Met, was rapidly phosphorylated in the nemosis-stimulated keratinocytes. Nemosis-induced in vitro scratch-wound healing was inhibited by an HGF-neutralizing antibody as well as the small molecule c-Met inhibitor, SU11274. HGF-induced in vitro scratch-wound healing was inhibited by PI3K inhibitors, wortmannin and LY294002, while LY303511, an inactive structural analogue of LY294002, had no effect. Inhibitors of the mitogen-activated protein kinases MEK/ERK1/2 (PD98059 and U0126), and p38 (SB203580) attenuated HGF-induced keratinocyte in vitro scratch-wound healing. We conclude that nemosis of BMSCs can induce keratinocyte in vitro scratch-wound healing, and that in this effect signaling via HGF/c-Met is involved.
我们之前表明，人真皮成纤维细胞间的细胞接触可在一个被称为“细胞程序性坏死”（nemosis）的过程中诱导肝细胞生长因子/分散因子（HGF）的表达。现在我们报告骨髓间充质干细胞（BMSC）中细胞程序性坏死的起始情况。由于BMSC在细胞移植治疗中的应用越来越广泛，我们旨在证明BMSC细胞程序性坏死对伤口愈合的功能作用和益处。在划痕伤口愈合试验中，使用细胞程序性坏死的细胞和单层细胞来刺激HaCaT角质形成细胞迁移。BMSC球体中诱导了细胞程序性坏死的两个指标，即HGF产生和环氧合酶-2表达。与等量的单层细胞相比，细胞程序性坏死的细胞以浓度依赖的方式在体外诱导角质形成细胞划痕伤口愈合。HGF受体c-Met在细胞程序性坏死刺激的角质形成细胞中迅速磷酸化。细胞程序性坏死诱导的体外划痕伤口愈合受到HGF中和抗体以及小分子c-Met抑制剂SU11274的抑制。HGF诱导的体外划痕伤口愈合受到PI3K抑制剂渥曼青霉素和LY294002的抑制，而LY294002的无活性结构类似物LY303511则没有作用。丝裂原活化蛋白激酶MEK/ERK1/2（PD98059和U0126）和p38（SB20358）的抑制剂减弱了HGF诱导的角质形成细胞体外划痕伤口愈合。我们得出结论，BMSC的细胞程序性坏死可诱导角质形成细胞体外划痕伤口愈合，并且在这种作用中涉及通过HGF/c-Met的信号传导。
