title: IKK-2 is required for TNF-alpha-induced invasion and proliferation of human mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) can contribute to tissue repair by actively migrating to sites of tissue injury. However, the cellular and molecular mechanisms of MSC recruitment are largely unknown. The nuclear factor (NF)-kappaB pathway plays a pivotal role in regulating genes that influence cell migration, cell differentiation, inflammation, and proliferation. One of the major cytokines released at sites of injury is tumor necrosis factor-alpha (TNF-alpha), which is known to be a key regulator of the NF-kappaB pathway. Therefore, we hypothesized that TNF-alpha may lead to MSC invasion and proliferation by activation of the NF-kappaB pathway. TNF-receptor 1 and 2, NF-kappaB (p65), and IkappaB kinase 2 (IKK-2) are expressed in human MSCs (hMSCs). Stimulation of hMSCs with TNF-alpha caused a p65 translocation from the cytoplasm to nucleoplasm but did not change the expression profile of MSC markers. TNF-alpha strongly augmented the migration of hMSCs through the human extracellular matrix. Using lentiviral gene transfer, overexpressing a dominant-negative mutant of IKK-2 (dn-IKK-2) significantly blocked this effect. NF-kappaB target genes associated with migration (vascular cell adhesion molecule-1, CD44, and matrix metalloproteinase 9) were upregulated by TNF-alpha stimulation and blocked by dn-IKK-2. Moreover, using the bromodeoxyuridine assay, we showed that the inhibition of the NF-kappaB pathway caused a significant reduction in the basal proliferation rate. TNF-alpha stimulated the proliferation of hMSCs, whereas overexpression of dn-IKK-2 significantly blocked this effect. TNF-alpha led to the upregulated expression of the proliferation-associated gene cyclin D1. In conclusion, we demonstrated that the NF-kappaB pathway components, p65 and IKK-2, are expressed in hMSCs. Our data provide evidence that this signal transduction pathway is implicated in TNF-alpha-mediated invasion and proliferation of hMSCs. Therefore, hMSC recruitment to sites of tissue injury may, at least in part, be regulated by the NF-kappaB signal transduction pathway.
authors.affiliation: Experimental Surgery and Regenerative Medicine, Department of Surgery, University of Munich (LMU), Nussbaumstrasse 20, 80336, Munich, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Böcker
authors.forename: Denitsa
authors.initials: D
authors.lastname: Docheva
authors.forename: Wolf Christian
authors.initials: WC
authors.lastname: Prall
authors.forename: Virginia
authors.initials: V
authors.lastname: Egea
authors.forename: Emmanouil
authors.initials: E
authors.lastname: Pappou
authors.forename: Oliver
authors.initials: O
authors.lastname: Rossmann
authors.forename: Cvetan
authors.initials: C
authors.lastname: Popov
authors.forename: Wolf
authors.initials: W
authors.lastname: Mutschler
authors.forename: Christian
authors.initials: C
authors.lastname: Ries
authors.forename: Matthias
authors.initials: M
authors.lastname: Schieker
mesh_terms: D015720:5'-Nucleotidase / Q000032:analysis; D015703:Antigens, CD / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects; D001692:Biological Transport / Q000187:drug effects; D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D000071063:Endoglin; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000032:analysis; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D018800:Thy-1 Antigens / Q000032:analysis; D051996:Transcription Factor RelA / Q000378:metabolism; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D011956:Receptors, Cell Surface; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D018800:Thy-1 Antigens; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D051550:I-kappa B Kinase; D015720:5'-Nucleotidase