Overexpression of dnIKK in mesenchymal stem cells leads to increased migration and decreased invasion upon TNFα stimulation
•HMSC were successfully transduced with a dominant-negative mutant of IκB kinase 2.•Characterization of dnIkk MSC showed unaltered expression of TNFα receptors.•3-D invasion towards the TNFα was significantly reduced after blocking IKK-2.•2-D migration assay revealed a strong migratory response of h...
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Published in | Biochemical and biophysical research communications Vol. 436; no. 2; pp. 265 - 270 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
28.06.2013
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Subjects | |
Online Access | Get full text |
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Summary: | •HMSC were successfully transduced with a dominant-negative mutant of IκB kinase 2.•Characterization of dnIkk MSC showed unaltered expression of TNFα receptors.•3-D invasion towards the TNFα was significantly reduced after blocking IKK-2.•2-D migration assay revealed a strong migratory response of hMSC towards TNFα.•Blocking of IKK-2 resulted in a significantly increased 2-D migration of hMSC.
IκB kinase 2 (IKK-2) mediates tumor necrosis-factor α (TNFα) induced invasion of human mesenchymal stem cell (hMSC) to sites of tissue injury. Suppressing IKK-2 activity leads to reduced expression of proteolytic enzymes and impaired invasive capacity. In order to further reveal mechanisms of hMSC recruitment, we here aimed to analyse the impact of IKK-2 on two-dimensional migration upon TNFα stimulation in contrast to three-dimensional invasion. An immortalized hMSC line (SCP-1) was transduced with a dominant-negative mutant of IκB kinase 2 (SCP-1 dnIKK). Migration was assessed using a linear-gradient chemotaxis chambers by time-lapse analysis. Invasive capacity through human extracellular matrix was analysed using transwell invasion assays. RT-PCR confirmed increased IKK-2 expression levels in SCP-1 dnIKK cells, while TNFα receptor I and II expression was not altered. Invasion upon TNFα stimulation was significantly reduced by 78% in SCP-1 dnIKK. In contrast, migration was significantly increased, represented by a 60% elevated forward migration index and a 2.1-fold higher mean dislocation of the center of mass towards TNFα. In conclusion, our data confirms the impact of IKK-2 in TNFα dependent hMSC recruitment. Interestingly, reducing IKK-2 function increases two-dimensional migration towards TNFα, while invasive capacity is impaired. These findings contribute to a deeper understanding of MSC’s biological properties orchestrating the complex processes of stem cell recruitment and homing. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0006-291X 1090-2104 |
DOI: | 10.1016/j.bbrc.2013.05.091 |